Past Activities 2017-2018

POSTED ON BEHALF OF CASPER LINNESTAD
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Dear participants of the Online Forum on Synthetic Biology,

My name is Casper Linnestad and I work in the Norwegian Ministry of Climate and Environment. I am the national focal point for the Cartagena Protocol on Biosafety. Furthermore, I was a member of the Ad Hoc Technical Expert Group on Synthetic Biology that met in Montreal in 2015. My background is within genetics and gene technology and the regulation of LMOs.

At the last COP in Mexico (in decision XIII/17) the mandate of the AHTEG on synthetic biology was extended with new terms of reference. The open-ended online forum was also extended to support the work of the AHTEG. The online discussions ahead of us on synthetic biology will constitute an important basis for further work of the AHTEG.

As you know, the discussions will last for several months, ending in October. For clarity and structural purposes, the Secretariat has divided the online discussions in different topics. The topic of our first discussion (to be held from July 3rd to 17th) is directly linked to the terms of reference of AHTEG:

“Review recent technological developments within the field of synthetic biology to assess if the developments could lead to impacts on biodiversity and the three objectives of the Convention, including unexpected and significant impacts”.

In the first topic of the online discussion I would like to invite the participants to focus the discussion specifically on how developments within the field of synthetic biology could lead to impacts on biodiversity in the context of the three CBD objectives, namely (i) the conservation of biological diversity, (ii) the sustainable use of its components and (iii) the fair and equitable sharing of the benefits arising out of the utilization of genetic resources.

Please note that several submissions relating to this topic have already been made by Parties and others following an invitation by the Secretariat, to be found at http://bch.cbd.int/synbio/submissions/2017-2018.shtml. Several of these submissions outlined the availability of techniques that are being commonly used in synthetic biology, such as CRISPR and TALENs. Some of the submissions also listed applications in which these techniques are used, including biofuel production, bioremediation and mosquito population control.

In order to focus the discussion over the next two weeks, I would like to suggest that we concentrate on the following three guiding questions:

1) What are the potential negative impacts, including unexpected and significant adverse effects, of the most recent technological developments in synthetic biology on biodiversity and the three objectives of the Convention?

2) What research and cooperation activities are being conducted on the possible benefits and potential adverse effects of organisms, components and products of synthetic biology on biodiversity to fill knowledge gaps and identify how those effects relate to the objectives of the Convention and its Protocols?

3) Are there other recent technological developments that have taken place within the field of synthetic biology that need to be considered in this discussion?

When sharing your views, I kindly ask you to be concise and focused. Whenever possible, please avoid lengthy introductions and unnecessary repeating of previous posts. I am very aware of the fact that you are all very committed to other tasks. Nevertheless, I sincerely hope that you will be able to participate in this forum and trust that it will be informative, constructive and beneficial to all of us.

It is an honor for me to moderate this discussion and I am looking forward to an inspiring an lively debate.

Best wishes and good luck!
Casper Linnestad

Dear all,

- What are the potential negative impacts, including unexpected and significant adverse effects, of the most recent technological developments in synthetic biology on biodiversity and the three objectives of the Convention?

First one, I considers that the “operational definition” must be used and shared in others fora to be enriched too, i.e. WTO, WIPO and Liaison Group of the Biodiversity-related Conventions. In my perspective, there isn’t doubt that the definition is a part of the scope of the CBD system (lege data) in a way that the Nagoya Protocol is the only one instrument, for the moment, to regulate this topic.

Second one, in my legal opinion, the CBD and the Nagoya Protocol are international programmatic regimes that will regulate future aspects in biotechnology (lege ferenda), “operational definition” is included, in accordance with the objective and purpose of both treaties.

Finally, the potential negative impacts are legal gaps (limbo) and fraud derives from choice of jurisdiction fora between Contracting Party and Non-Contracting Party of the CBD and the Nagoya Protocol. This negative impacts could provokes erosion to both international regimens in favor to private negations of industry (i.e. pharmaceuticals and chemicals industries).

POSTED ON BEHALF OF ANGELA LOZAN
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Hello,

I would like to thank the colleagues for interesting examples discussed in the forum. I would support them and wish to listing some others possible negative effects from SynBioon ecosystems and biodiversity that can be taken in consideration.

Ecosystem conservation:
Biofuel and bioenergy on SynBio: biomass extraction from existing agricultural practices can leading to decline in soil fertility and structure. Removing corn plantsfrom fields would require significant additional use of nitrogen, phosphorous and potassium fertilizers.

An environmental harm can be caused by establishing plantations in former forests, harvesting natural grasslands, pressures on deserts and wetlands.

Biosafety considerations related to the accidental or intentional release of SynBio organisms  and the failure to be due to the unpredictable complexity of natural microbial ecosystems.

Xenobiology.  Acknowledge the possibility of invasiveness and unintended effects from xenobiology.  

There is also concern about indirect impacts of the promises of synthetic biology and de-extinction. 

“In-situ” conservation:
Support for in situ conservation may decrease with the expectation that lost species will be able to be resurrected

The transfer of genetic material from an organism resulting from synthetic biology techniques to another organism would change biodiversity at a genetic level (genotype) and could spread undesirable traits (phenotype).

The organisms resulting from synthetic biology techniques becoming invasive or disrupting food chains or through the transfer of DNA from sexual gene flow or horizontal gene transfer. 

Bio-plastics:
Some methods of producing biodegradable plastics may have more environmental impacts such as carcinogens and eutrophication than fossil-based polymers.

AL

With regard to ecosystem conservation, especially biofuel and bioenergy extraction from existing agricultural practices, of sure, removing corn, sorghum or millet plants from farms (fields) exposes soil structure to erosion. Hence, it will require significant additional use of fertilizers (nitrogen, phosphorus, potassium). This is the case of many Countries from West and Central Africa (including Cameroon where I am from). This is a great issue to address. In fact, in most of these countries, sorghum and millet rods are entirely used as firewood (or fuelwood), due to insufficient woody species around the huts.
Hence, how can synthetic biology be useful to create a novel product for biofuel and bioenergy? Although it is well known that replacing natural products with synthetic ones could reduce pressure on natural habitats, it could also disrupt conservation projects and displace small-scale farmers.

One category of issues raised relates to socio-economic changes consequent on the introduction of new synbio products or processes.  However these issues do not seem different in nature from those of any new technology, whether incremental or disruptive.  This seems nicely illustrated by Ms Therese [#8416] - use of biomass as feedstock for (hypothetical) large-scale biofuel production (synbio-based or otherwise) could have indirect effects on fertiliser use, soil quality etc as she indicates.  However she points out that this is already an issue, with agricultural biomass already being used for fuel by the very ancient method of burning it as firewood. 

This is not to argue that there are no issues in this regard, rather that such issues are not new or unique to synbio.  Consequently, they probably do not need new or unique regulation or provision.

The same applies to indirect ecological effects acting through this mechanism, also illustrated by  Ms Therese [#8416]: “Although it is well known that replacing natural products with synthetic ones could reduce pressure on natural habitats, it could also disrupt conservation projects and displace small-scale farmers”.  This issue would apply equally whether the synthetic product were derived from synbio, other novel manufacturing processes, or more traditional routes such as non-biological chemical synthesis, product substitution/replacement, etc.  So again, I would suggest these types of indirect effects, though potentially significant, are by no means new or unique to synbio and consequently probably do not need new or unique regulation or provision.  Indeed to do so would risk developing an inappropriate regulatory landscape in which equivalent risks are treated very differently, i.e. regulation that is neither proportionate nor consistent.

Dear All,

My name is Jim Thomas - I work with ETC Group - a Civil society organization that monitors developments in emerging technologies (including Synthetic Biology) and in the past few years I had the privilege  to serve on the AHTEG on Synthetic Biology as well as attend the relevant discussions at COP and SBSTTA. I commend Casper Linnestad on opening up this discussion so clearly and strongly urge fellow forum participants that we should keep to the task at hand which is to "Review recent technological developments within the field of synthetic biology to assess if the developments could lead to impacts on biodiversity and the three objectives of the Convention, including unexpected and significant impacts”.

With that charge in mind I particularly appreciated Barbara Livoreil’s contribution as an attempt to lay out what are those "recent technological developments  within the field of Synthetic Biology”  and strongly agree with her high level  list of some recent developments in the field  :

1/ That there is a move from a focus on microorganisms  to all living beings, plants, animals (and indeed insects as is apparent by the high representation of syn Bio insect-folks on this forum (Mssrs Alphey, James, Bier, Target malaria etc)
2/ That there is a move beyond the lab and containment (that has been eloquently echoed by  Taye Birhanu, Ed Hammond and and others raising concern about containment questions)
3/ That the hype of ‘precision' is foundering on the reality  of off-target and unexpected changes  at the genomic level as a result of new techniques - and while Anthony James may say its too early to make general statements about off-target effects pointing to as yet undeveloped techniques.. what is clear is it is certainly  inappropriate to make general statements about precision or predictability.
4/ That there are rising attempts to apply these techniques  to overcome the  principles of natural evolution (e.g. by developments in xenobiology, or by Mutagenic Chain Reaction/gene drives overcoming mendellian inheritance etc)

I also appreciate Ms Jashima Yasin’s  additions that at the applications level we are seeing a move to food and medical applications - that the applications are moving firmly beyond initial biofuels/biomaterials applications that characterized the field.

If I could suggest a clarifying schema , it might be useful to parse “recent technological developments within the field of Synthetic Biology” at the following levels:

Developments in Tools and Techniques
Developments in Applications
Developments in Systems, Platforms and approaches.

and then we can more systematically start to answer some of Casper’s more pointed  initial questions that involve actual assessment.  (since we are charged to actually “assess”  if the developments could lead to impacts on biodiversity and the three objectives of the Convention, including unexpected and significant impacts.

So below are some  reflections in this direction. I apologise in advance that its a long intervention.

Recent technological developments in synthetic biology tools and techniques.

a) Expanded Gene Editing techniques.
While Casper was right to highlight the most common gene editing techniques (CRISPR-CAS9 and TALENS in his introduction) we are also seeing a diversity of new gene editing tools emerge. For example Fang Zheng of the Broad institute has described four new CRISPR associated proteins that can operate similar to the CAS9 system including Cpf1, There is also a mini-Cas9 system taken from  Staphylococcus aureus.  Synthetic Genomics and The Venter Institute recently announced a new bacterial genome editing system that combines CRISPR Cas9 with a yeast based system. Novozymes recently entered into an agreement with Hebei University to use Natronobacterium gregoryi Argonaute (NgAgo) as a DNA- guided endonuclease for gene editing that would be a different approach from CRISR/TALENS/ZFN etc. Work by the Church lab in Harvard has recently used Lambda Red to edit bacterial genomes etc etc. I’m sure there’s more. When Dr James refers to attempts to reduce off-target effects from CRISPR that also speaks to ongoing developments in the underlying tools and techniques (in the meantime however the use of CRISPR-CAS9 is clearly generating off-target effects as evidenced by several papers - e.g. examining off target effects in mice  and by the decision by prominent  synthetic biology teams to steer away from CRISPR specifically to avoid the problem of off-target effects).

Do these new editing techniques impact biodiversity and the aims of the convention? yes in as far  as they may give rise to unpredictable and unexpected changes in the genome sequence and thereby in the phenotype of an organism or its offspring that changes the risk assessment . We have seen how small single point deletions and changes can have significant unexpected changes in an organism’s functioning and in the expression of other sequences so there is reason to be wary of undetected off-target changes. Until such a time as it can be shown beyond reasonable doubt that a new ‘editing’ approach is entirely predictable with no off-target effects elsewhere on the genome or epigenome , it would be precautionary  to treat genome editing as highly experimental and not ready for commercial or  ecological release.

b) improved DNA synthesis - It is my impression that the speed ,lowered costs and growing length of sequences for synthetic DNA construction is moving ahead at a pace and that recent high profile initiatives such as the HGP-write project sponsored by Autodesk and others is intending to further accelerate these trends. The implication is that synthesis may overtake gene editing as the quicker and more economical way to generate novel changes in a genome - and indeed, along with gibson assembly and other methods, may mean that whole genome synthesis could soon become more common for shorter genomes. Others who are following synthesis technology more closely can tell me if my speculation is off-base here.

Does rapidly improved, cheaper, faster DNA synthesis impact biodiversity and the aims of the convention? Yes, along with more rapid gene editing and new robotic/AI construction platforms (see below) it reduces  the barriers to fabricating novel organisms which means we are already seeing  a rise in the quantity and diversity of engineered organisms being produced both in labs and for commercial use. It also means that far more novel genomic arrangements are economically possible - widening the gap between what is known from nature as a starting reference point for biosafety assessments and what is entering the biosphere and commerce  by way of engineered organisms.  Also rapid, cheap synthesis as well as improvements in genome editing clearly challenge the access and benefit sharing arrangements since they lower the barriers to ‘digital biopiracy’ and increase the waterfront across which states will need to carry out surveillance for potential biopiracy.

c) The application of big data, machine learning, Artificial Intelligence and robotics to genome construction.
In my view possibly the most significant set of developments in the field of synthetic biology relates to the application of big data, machine learning algorithms (AI) and high throughput  robotic construction techniques to generating novel bioengineered organisms. This convergence is promised to  enable rapid and automatic prototyping of highly novel organisms. Here are 2 examples of what I mean:  Zymergen is a US synthetic Biology company that applies high throughput robotic construction and Artificial intelligence (machine Learning) to rapidly design and optimise novel microbial strains. In effect clients specify the desired out puts from a novel microbe (e.g. novel chemicals or natural product compounds) and the robots and AI’s collaborate to develop the industrially optimized genome sequence for that output. Bloomberg described Zymergen’s method this way:  “Zymergen’s algorithms suggest making 1,000 or so changes to the microbe’s genetic material . Then the robots take over, injecting the suggested DNA snippets into the specimens, testing their properties, collecting data and feeding that information back into the data trove.”. Zymergen’s highly automated massively parallel robotic labs are a far cry from the image of a careful  lab bench genetic engineer developing single new bio-inventions at human speed. 2) Amyris Biotechnologies is commercializing the work developed by DARPA, the US Defence Advanced Research Projects Agency, to create 'living foundries’. Similarly robotized as Zymergen, Amyris offers the ‘pathways programme’ promising commercial partners that  “Through the Pathways Program, partners can, with a small initial investment,  sponsor and secure a molecule” In effect, name a molecule for biosynthetic production and then let the automated high throughput processes attempt to design the microbe that will manufacture it.    Underlying these enterprises are rapidly expanding genomic databases such as Genbank and divseek (it is estimated that by 2025 genomic databases will contain 1 zetabase of genome sequencing data (1 thousand million trillion Base Pairs) encompass  2.5 million plant and animal genome sequences  and include genome sequences of all 1.2 million known species). While those sizes of datasets are dizzying for the human brain to comprehend  they are perfect fodder for mining by AI/machine learning for robotic construction set-ups.

What does this mean for biodiversity and the 3 aims of the convention?: We may increasingly see organisms that are not designed in any way a human being can easily understand but rather as a result of complex AI design processes. Carrying out biosafety assessments will become more challenging than it already is since the path and rationale behind the introduced changes will be opaque at best. . At this point the question of containment will become extremely critical (if its not already!) Automation and Ai in genome construction and ebioengineering  may also mean we can expect a large increase in the numbers of novel organisms and the speed with which ever greater novelty hits the market and the environment. Our current biosafety systems are designed for ‘case by case’ assessment of novel organisms arriving at a relatively slow rate at the assessors door and then considered by ‘expert’ human beings at a a human pace. Once novel genomic inventions are being churned out by automated AI-driven robots at cheaper and quicker rates that may overwhelm biosafety offices  capacity to assess and regulate. Does that then usher in AI-driven biosafety programmes to respond?!? How confident are we to hand both the rapid redesign of nature and the job of policing biosafety to AI’s and algorithmic cops that may ultimately prove flawed or beyond human comprehension. This may sound sci-fi right now but I think its the direction that these underlying developments in the field of synthetic biology take us.

Recent technological developments in synthetic biology applications.

This is of course a much wider field and having already written at length on tools and techniques i’d highlight just a few that seem relevant to the CBD context:

a) Mutagenic Chain Reaction (Gene Drives) and “active genetics” - It is clear from the submissions, comments to date and the specific individuals who have joined this conversation that the online forum, AHTEG, SBSTTA and COP will likely spend considerable time addressing and debating  MCR/Gene Drives in the coming months and years. the relevance of these applications to conservation of biodiversity is clear to all both on the claimed benefits (invasive species/ health/agronomic benefits) and the significant risks in terms of impacts on populations, ecosystems, biodiversity, agronomic changes and socio-economic and military threats. Through this process there needs to be a clear probing of assumptions and claims. Will MCR/gene drives even work?. What will be the ecological impact of resistance developing to MCR/gene drives? How might off-target effects arising from use of CRISPR constructs in these systems persist and propagate and to what effect? what range of gene drives are we talking about (not just CRISPR drives?) Will resistance be used to claim that MCR/Gene Drives can be localized and controlled (as opposed to global gene drives) and if so how will that impact potential adoption - e.g. in Agriculture. Can local gene drives inadvertently become global drives? Is the idea of “local drives” real or just an aspiration? While significant philanthropic funds from a few sources have been targeted into framing MCR/Gene Drives through one or two optimum use cases (malaria, island rodent extinction, schistosomiasis) the real commercial vale of MCR/Gene Drives (especially so called "local drives") will be in agriculture so it will be important to consider agricultural use cases (e.g. agricultural pest eradication, reversing herbicide resistance in weeds, introducing new herbicide susceptibilities) and the likely biodiversity impacts of agronomic changes proceeding from the uses. I notice Dr Bier of UCSD has joined this online forum. Dr Bier is pioneering the field of ‘active genetics’ and it would be helpful if he and his colleagues could share information about what other applications they see in the ‘active genetics’ domain beyond the Mutagenic Chain Reaction.

b) Development of synthetic microorganisms for agricultural use. - Specifically we are seeing work on  synthetically engineering micro-organisms for nitrogen fixation in the roots of crops, Synthetically engineering organisms to increase carbon sequestration in soils, synthetically engineering endophytes to ‘prime’ plants for resistance traits such as herbicide resistance or abiotic stresses, synthetically engineered organisms to deliver growth hormones to plant roots, synthetic microorganisms that form biofilms intended as ‘crop protection' and synthetic microorganisms for altering signalling between plants and  rhizome communities and for soil remediation.  Soil ecology and soil microbial biodiversity  is both particularly complex and still  poorly understood and so applications that either set out deliberately or inadvertently  alter soil microbial populations may require particularly careful assessment and high degree of precaution.

c) Ecosystem-level engineering by Synthetic biology -  The US Defense Advanced Research Project Agency (DARPA) - the  government funding agency currently providing the largest share of funds for synthetic biology research - is allocating priority funds to  researchers bioengineering insects to carry  engineered plant viruses to agricultural crops as a targeted gene therapy approach for agriculture. DARPA explains this so-called ‘insect allies’  approach like this: “Insects eat plants and insects transmit the majority of plant viruses… DARPA plans to harness the power of this natural system by engineering genes inside plant viruses that can be transmitted by insects to confer protective traits to the target plants they feed upon.” DARPA is additionally funding researchers to re-engineer the ecological niche preferences of organisms (that is developing technologies that enable the genetic engineering of an organism’s preference for a niche (e.g., temperature range, food source, and habitat) while also funding work to “reprogramme larval behaviour at sea’ which aims to develop "genetic, epigenetic, biochemical, microbial, or conditioning-based approaches that alter settlement habitat preferences in a lab-raised cohort of larvae". Like MCR/Gene Drives and ‘active genetics’ these systems which are ultimately  for environmental release deliberately aim at altering biodiversity and population structures at an ecosystem level as well as the genetic level. Along with MCR/Gene Drives and "active genetics" this seems to signal a further shift to synthetic biology being used not only to alter genomes but also to deliberately alter whole ecosystems and populations via genetic engineering as the point of leverage - namely ecosystem engineering.

d)  - Development of methanotroph based biosynthesis platforms  - A growing number of synthetic biology companies (e.g. Calysta, Intrexon, Coskata) as well as government research programmes are ‘reprogramming methanotrophs to convert natural gas and other fossil carbon sources to high value fuels, chemicals and consumer compounds. These platforms shift the economics of fossil fuel extraction  by offering the option to more easily  upgrade low value fossil fuels into high value commodities. The biodiversity costs of fossil fuel extraction (including for methane via fracking, coal bed methane, methane hydrates) are well documented. These synthetic biology developments therefore are alarming for potentially extending and deepening the value of fossil fuel extraction activities. Much of the discussion of Synthetic biology to date (framed around initial ‘bioeconomy/biomass’ uses ) have come with hyperbolic claims of moving economies past fossil fuel dependencies but these developments, by contrast, seem to be deepening those dependencies.

  3 - Recent technological developments in synthetic biology systems, platforms and approaches

An example of en emerging platform development relevant to this topic and the mandate of the CBD   is the emerging field of 'Molecular Communication’ (MolCom). Molecular Communication describes the application of information theory to molecular  processes where communication signals (e.g. between organisms) are physically encoded in molecules (rather than by waves) - This includes biological signalling processes  (such as pheromones, hormones) but also genetic and epigenetic communication over space and time eg via DNA, RNA. Within the field of mol com there is a range of applied work on  deliberately interfering and intervening in molecular signalling processes and trying to harness and redirect them - e.g. to carry messages between electromechanical and biological systems - such as using nano devices to release molecular signals that effect genetic or epigenetic changes in an organism. MolCom researchers talk of developing an "internet of nanobio-things” where the digital world can talk with the biological. Work on artificially inscribing data into biological molecules - e.g. using DNA as a storage medium or encryption medium - also falls into this category. In my view developments in RNAi can also be seen from a molCom perspective since small synthetic messenger molecules are being emitted as genetic signals (either as sprays or from engineered plants) with the intention of disrupting an existing ‘communication’ process (gene expression). Synthetic microbes that aim to alter molecular signalling between soil microbes and plant roots (e.g. to stimulate growth) are also effectively ‘mol com’ technologies as are DARPA’s ‘insect allies’ project which harnesses insects as a carrier for a signal inscribed in a viral carrier.

I realize that this is a very long intervention - i hope however  it helps answer the task we were assigned to "Review recent technological developments within the field of synthetic biology” and  to begin to assess them.

Dear all, dear Casper,

It is a pleasure to contribute to this online forum. As requested by Casper, we like to focus on the specific questions posed.

1) What are the potential negative impacts, including unexpected and significant adverse effects, of the most recent technological developments in synthetic biology on biodiversity and the three objectives of the Convention?

We would have preferred if this question would mention the potential for both positive and negative impacts of the most recent technological developments in synthetic biology as well as of their applications. On a case-by-case basis these technological developments and their applications can have positive, neutral of negative effects on biodiversity and the three objectives of the Convention. This topic has been iterated in depth in both the previous on-line discussion as well as the AHTEG, and we consider this topic does not need further deliberation.

2) What research and cooperation activities are being conducted on the possible benefits and potential adverse effects of organisms, components and products of synthetic biology on biodiversity to fill knowledge gaps and identify how those effects relate to the objectives of the Convention and its Protocols?

In the Netherlands the following activities have been and are currently undertaken.
•Four reports commissioned by the Dutch National Institute for Public Health and the Environment (RIVM) were delivered. They describe experience gained with environmental risk assessment of LMO’s and new developments in white, green and red biotechnology. These RIVM reports van be can be found on:  http://www.stw.nl/nl/content/biotechnology-and-safety.
•Based on these reports a call for research into the safety aspects of these new developments in modern biotechnology - including synthetic biology - was released last year.
•Ten different research projects have been awarded and will make a start this year. The overall budget of the program was 9 million Euro`s.
•Last year the RIVM issued a policy report on gene drives resulting in adjustment of the Dutch legislation for working with gene drives in contained use facilities (http://www.rivm.nl/Documenten_en_publicaties/Wetenschappelijk/Rapporten/2016/februari/Gene_drives_Policy_report). RIVM is now working on aspects of risk assessment for contained use of gene drives in cooperation with several European partners.
•Modern biotechnology - including synthetic biology - is developing and maturing rapidly. At RIVM the potential impact of these new developments on risk assessment methodology is currently being researched. A policy report is expected towards the end of 2017.
•The Commission of Genetic Modification, together with the Health Council of the Netherlands, published in 2016 a report that describes major new developments and applications in biotechnology and possible stumbling blocks and (ethical and societal) dilemmas which arise from these developments and trends. This trend analysis can be found on:
http://www.cogem.net/index.cfm/en/publications/publication/trend-analysis-biotechnology-2016

3) Are there other recent technological developments that have taken place within the field of synthetic biology that need to be considered in this discussion?

•The application of CRISPR Cas-based gene drives in organisms, especially in insects, needs to be considered with respect to potential benefits and potential risks on ecosystem level.
•The accessibility of the biotech tools (e.g. CRISPR) is increasing and is now also more or less available for the public (CRISPR kits can be ordered over the internet). This technology in general will become more and more accessible to e.g. DIY and other communities.
•Another new development is the development and application of external genome regulation methods, e.g. RNA interference in the form of sprays to control pests or influence plant characteristics, which raises questions about the risk assessment framework (if needed) for these applications.
•GM algae production platforms might be an important route for the production of chemical substances. The intrinsic need for relatively `open` (due to the need of sunlight) production ponds/facilities requires well designed safety and containment measures,  either physical of biological.
•Whole cell sensor development is being pursued more actively. Sensor use inside and outside the laboratory may require well designed containment strategies.
•Cell free systems - e.g. again for sensor type applications - are being developed. Risk assessment methodology for these type of applications needs to be scrutinized.
•An important element that needs to be stressed is the ever-increasing speed of development of the field of biotechnology. New biotechnology tools (e.g. CRISPR) combined with automated laboratories (e.g. `foundries`), DNA circuitry design tools and bioinformatics are important enablers for developing new techniques and applications. This development is supported by substantial research and investment funding.

Kind regards,
Jaco Westra and Boet Glandorf
GMO Office/Dept. of Gene Technology and Biosafety
Dutch National Institute for Public Health and the Environment (RIVM)

Dear all, dear Casper,

My name is Mr. Filemon N Shindume, working for the Ministry of Agriculture, Water and Forestry (Namibia) and had the privilege to serve on the AHTEG on Synthetic Biology as well as attended the relevant discussions at the last two COPs.  This is my first intervention in the forum so I would like to thank our moderator, and acknowledge with gratitude those who have posted before me.

I highly appreciate the contributions that attempt to lay out what are those "recent technological developments within the field of Synthetic Biology” including the citations/links provided.  Also, a long compiled intervention by Mr Jim Thomas [#8437] is very helpful to country with not so well developed civil society organizations.

Therefore, having a list I believe will accord us opportunity to assess case-by-case if the identified developments could lead to impacts on biodiversity and the three objectives of the Convention, including unexpected and significant impacts.

Thank you,

Dear participants, thank you for being so active and presenting a wide array of interesting views! I am very grateful for this. No doubt, your efforts will be much appreciated in subsequent rounds of discussions and work under the CBD.

We are nearing the midpoint of the discussion and I would like to reiterate that the topic of our discussion is how developments within the field of synthetic biology could lead to impacts on biodiversity in the context of the three CBD objectives (the conservation of biological diversity, the sustainable use of its components and the fair and equitable sharing of the benefits arising out of the utilization of genetic resources).

There are many ways to address this topic. Some of you have chosen to make use of the guiding questions that I posted on Monday:

1) What are the potential negative impacts, including unexpected and significant adverse effects, of the most recent technological developments in synthetic biology on biodiversity and the three objectives of the Convention?

2) What research and cooperation activities are being conducted on the possible benefits and potential adverse effects of organisms, components and products of synthetic biology on biodiversity to fill knowledge gaps and identify how those effects relate to the objectives of the Convention and its Protocols?

3) Are there other recent technological developments that have taken place within the field of synthetic biology that need to be considered in this discussion?

For others, it was important earlier this week to bring to the table the current operational definition and trying to improve it, as a first step. Although having sympathy for this view (it is always worthwhile to improve!), I thought we were better off concentrating on the issues mentioned above, as I find the definition already quite inclusive and flexible.

We have the benefit of drawing upon the extensive experience of the Secretariat, which very wisely has advised us to divide the online discussions into different topics. In the remaining days to follow, I ask everyone to stick to our task and topic.

Lastly, a quick remark to the suggestion today from Jaco Westra and Boet Glandorf that guiding question 1) could also capture positive impacts of synbio organisms on biodiversity. I see no harm in this, it simply adds to the picture!

Keep up the good work and let the sentences flow!
Sincerely,
Casper

To the four fallacies in defense of the AHTEG definition [#8393], I should now add special pleading  [#8442].  Contrary to assertions that the AHTEG defintion is “inclusive” and “flexible” are its words: “modification of genetic materials, living organisms and biological systems”.  An example may elucidate both exclusion and inflexibility: The Naked-Mole Rat Genome Resource (http://naked-mole-rat.org) . Quoting from the “new and emerging issue” for COPXIV submitted by The Peruvian Society of Environmental Law:  “The ‘BLAST[ing]’ of the mole-rat in 2014 enabled scientists to conduct R&D without ever having to handle any genetic material or even having to identify themselves as working on the genome. According to The Naked Mole-Rat Genome Resource: ‘There are no restrictions on the use of our portal or the genome data.’” (Peruvian Society of Environmental Law,  2017, p. 3). Eight patents have been granted on the value added and thirty more are pending.

The Naked-Mole Rat Genome Resource and similar endeavors would not meet the AHTEG definition of synthetic biology nor would they incur an ABS obligation which applies to “genetic material”.

Peruvian Society of Environmental Law (SPDA), (2017). “Lawful Avoidance of ABS: Jurisdiction Shopping and Selection of non-Genetic-Material Media for Transmission”. In response to “Proposals for new and emerging issues for SBSTTA-21 and COP-14.  (SCBD/OIC/DC/RH/84326).  In English:  https://www.cbd.int/doc/emerging-issues/SPDA-submission2017-05-en.pdf  or in Spanish: https://www.cbd.int/doc/emerging-issues/SPDA-submission2017-05-es.pdf

Dear all, dear Casper Linnestad,

I am Silvia Ribeiro, Latin America Director for ETC Group. I have been participating in the synbio forum and all related discussions at SBSTTA and CBD.

Picking up on the question of reviewing recent technological developments in the field of synthetic biology and also the observation by Ms Jashima Yasin that at the applications level we are seeing an increasing move to food and applications applied to the human body,  I would like to draw your attention to a resource developed by ETC Group that is currently in beta phase but may be useful for this discussion:

The GMO 2.0 Ingredients database is hosted at http://database.synbiowatch.org. This is a searchable database  of synthetic biology derived ingredients (that is ‘ products of synthetic biology’ in CBD parlance). It includes ingredients that are already on the market, that are close to the market, or that are under development. The information here was compiled through desk research, and will be updated as new ingredients or information emerges so is necessarily partial however it indicates the scope of compounds, including natural product replacements, now being developed from the application of synthetic biology. The database holds 70 entries for syn-bio derived compounds thought to already be on the commercial market, 72 entries for syn-bio derived compounds ‘coming to market’ (where there is a commercial player involved and signalling intent to market) and 206 entries where we have not been able to ascertain how close the development is to commercial release.

The database helps illustrate for policymakers and the public that:

• There are now extensive efforts underway to replace natural product compounds with syn-bio derived versions. Examples include vanillin, resveratrol, asthaxantin, anthocyanins, steviol rebausides, shea and cocoa butter replacements, betaine, citrus oils, beta elemene, patchouli, rose scent, Lactones, saffron

• These products of synthetic biology are now going into foods, cosmetics, supplements, probiotics, essential oils and other markets

ETC Group have additionally helped prepare an online searchable map showing the countries whose agricultural economies and farmers livelihoods may be affected by the issue of synbio replacements for just 13 of these natural products (it does not extend to the full 350 or so entries from the database!).

The Natural Products Map is here: http://www.synbiowatch.org/commodities/natural-products-map/

This speaks directly to the second aim of the convention - the impact on sustainable use of biodiversity since replacements can alter the market and therefore support for sustainable use of products of biological diversity and biodiversity-based livelihoods.

I am Tom Nickson, an independent science policy consultant, with more than 20 years of experience with environmental risk assessment for LMO’s and 13 years of experience with the CBD and its protocols.  I begin this intervention by thanking the Secretariat of the CBD for providing this form and the moderator for providing a framework.  I also thank those who have provided their ideas in a constructive and collegial manner. 

My intervention is a reaction to statements found in #8422 and #8437 that appear to declare unrealistic standards for environmental risk assessment.  Both statement appear to assert that the technologies under discussion are creating risks heretofore unknown to natural systems. 
#8422:  “Synthetic biology is again a tool just like any other breeding technique. A breeding technique should develop a new variety adding to biodiversity. It should not be deleterious to anything available in nature. No eradication except deadly pathogens.” 

#8437:  Until such a time as it can be shown beyond reasonable doubt that a new ‘editing’ approach is entirely predictable with no off-target effects elsewhere on the genome or epigenome , it would be precautionary  to treat genome editing as highly experimental and not ready for commercial or  ecological release.

In #8422, the second and third sentences apply a personal and arbitrary risk assessment standard to technologies that this forum may label as synthetic biology.  This statement may have been written in haste, but it overlooks the fact that a very large proportion of the world is managed to human-determined standards.  Conservation efforts manage ecosystems to achieve predetermined human values; never to a achieve an outcome of no deleterious effects to all biodiversity.  Similarly, regulatory decisions are informed using sound risk assessments that examine effects on beneficial biodiversity and recognize that the biodiversity characterized as “pests” e.g, invasive species, can have acceptable effects that control and even reduce their populations. 

Similarly, in #8437, many would argue that the approach suggested in not reasonable since it is primarily justified by speculation.  The precautionary approach demands evidence to justify the decision.  Furthermore, proper consideration must be given to the risk associated with severely limiting research especially when adequate protection may be afforded by risk management measures. 

Conversely. it is also worthwhile to highlight a statement concerning risk assessment made in #8427:  “The use of gene drive to either reduce the number of malaria vectors or to affect the ability of mosquitoes to transmit the disease is considered as one of the potential complementary tools to achieve malaria elimination. It does require responsible science and case-by-case risk assessment to ensure that the technology development takes into account stakeholders' concerns and that appropriate studies are carried-out in this direction.”  I believe that a dispassionate, scientifically-informed approach to technology evaluation as proposed here is prudent and appropriate. 

In closing, I thank those who have shared their technical, real-world expertise related to mosquito/malaria control (#8421 and #8430), and those who have provided us with many useful links (#8435 and #8439).  This information is valuable for informing this discussion.

Thanks you.

This is Paul Freemont and I had the privilege of being on the AHTEG group that met in 2015. I am a Professor and co-director of the Synthetic Biology Centre at Imperial College London.

I have very much enjoyed reading the posts so far and would like to contribute to the accumulating knowledge some interesting reports and commentaries.

Genome editing
UK government interim report https://www.publications.parliament.uk/pa/cm201617/cmselect/cmsctech/854/854.pdf

Nuffield council report on the bioethics of genome editing
http://nuffieldbioethics.org/project/genome-editing/ethical-review-published-september-2016

Response to the report by the Royal Society (UK academy of sciences) https://royalsociety.org/~/media/policy/Publications/2016/02-01-16-response-on-Bioethics-inquiry-into-genome-editing.pdf

European Academies' Science Advisory Council report on genome editing
https://www.knaw.nl/shared/resources/internationaal/bestanden/easac-report-31-on-genome-editing-20170323

Synthetic Biology as a tool for conservation and biodiversity
http://reviverestore.org/is-it-time-for-synthetic-biodiversity-conservation/

In terms of new technological capabilities i note a number of responses which seem to overstate the current technological capabilities (e.g. [#8437]). It is important to note that biotechnology and metabolic engineering are very developed fields with the aim of engineering microbial cell factories to produce material like chemicals or drugs using fermentation technology rather than using  synthetic chemistry. A main part of the synthetic biology field is to develop tools and new knowledge based approaches to accelerate and expand the use of fermentation technology for bio-manufacturing. This opens up possibilities of using waste streams for fermentation and also a distributed manufacturing opportunity where appropriate. The automation and knowledge based computational approaches is to make the design  process more efficient. It is worth noting that fermentation technologies are around 7,000 years old.

The imaginary future scenarios presented  as highly negative and impactful on biodiversity are pure speculation. It is impossible to predict the future applications of an emerging technology. However many threads rightly point out that regulatory and governances structures need to be adaptable to emerging applications and this is true for any new technology (e.g. AI and robotic humanoids). Also much more research is required to rigorously test the hypotheses that are posed by synthetic biology approaches in terms of release or accidental release. Also more research  funding is required to enable the regulatory agencies to develop tools that can be employed as part of the testing process.

In terms of biodiversity there are views within the professional conservation community which are suggesting that genomic technologies like genome editing and assembly could provide solutions to
current conservation problems (see reference above).

Personally I  would love to see an evidence based discussion which focuses on the challenges, risk and benefits of biotechnology to conservation and biodiversity as enabled by the new tools of synthetic biology - and gladly the forum is providing an excellent opportunity for such a discussion.

By way of a quick response to Mr Tom Nickson of Monsanto/CropLife/ American Seed Trade Association:

You state that "the precautionary approach demands evidence to justify the decision.  Furthermore, proper consideration must be given to the risk associated with severely limiting research especially when adequate protection may be afforded by risk management measures. "

I think the relevant use of precautionary approach here is that as stated by the Cartegena Protocol  (since we are talking about biosafety risks) . Article 10.6 and 11.8,  states "Lack of scientific certainty due to insufficient relevant scientific information and knowledge regarding the extent of the potential adverse effects of an LMO on biodiversity, taking into account risks to human health, shall not prevent a Party of import from taking a decision, as appropriate, with regard to the import of the LMO in question, in order to avoid or minimize such potential adverse effects."

I don't think the protocol directs parties to carry out a cost-benefit analysis in relation to impacts of a precautionary decision on research activities as you suggest above and in any case it is hard to see how deciding not to commercialize or environmentally release a gene edited organism in any way impacts disinterested scientific 'research' in the laboratory per se - what it impacts is commercial development (eg by your company) which is a different matter.

In any case, its not simple 'speculation' that CRISPR-CAS9 gene editing appears to give rise to off-target effects. I am aware that there is a concerted effort by companies invested in CRISPR to contain the implications of the recent mouse studies published in Nature methods showing unexpected mutations after CRISPR CAs9 use in vivo (for more on the commercial efforts by CRISPR companies  to discredit these studies see: http://www.gmwatch.org/en/news/latest-news/17714-gene-editing-companies-angry-at-research-revealing-unintended-crispr-effects)

However this question of concern about off target effects proceeds these studies - e.g.. as reported in Stat news last year its not helpful for biotechnology companies using CRISPR to have their "head in the sand" on the topic:  https://www.statnews.com/2016/07/18/crispr-off-target-effects/

Another mouse study published at the end of May in Nature Communications draws similar conclusions:  "CRISPR/Cas9 targeting events cause complex
deletions and insertions at 17 sites in the mouse
genome"  https://www.nature.com/articles/ncomms15464

Teh off-target effects are so clearly accepted that in response synthetic biologists are trying to design methodologies to reduce off target effects since " the potential for unanticipated downstream effects from off-target mutations is an important regulatory consideration for agricultural applications." - e.g. Wolt, J. D., Wang, K., Sashital, D., & Lawrence-Dill, C. J. (2016). Achieving Plant CRISPR Targeting that Limits Off-Target Effects. The Plant Genome, 9(3).
https://dl.sciencesocieties.org/publications/tpg/articles/0/0/plantgenome2016.05.0047

So this question of off-target effects from CRISPR is far from speculative, it would seem there is certainly enough evidence to invoke precaution by the standards of the protocol.

Interesting theres apparently enough evidence for CRISPR developers themselves to invoke precaution. In a recent paper from the Church Lab at Harvard "Design, synthesis, and testing toward a 57-codon genome", http://science.sciencemag.org/content/353/6301/819   the authors explained that thy chose not use CRISPR or the labs own MAGE approach for this ambitious undertaking exactly because of the off target effects of gene editing:  ""Although it is possible to simultaneously edit multiple alleles using MAGE or Cas9, these strategies would require extensive screening with numerous oligos and likely would introduce off-target mutations."  These are the scientists from the same institutions that further  developed CRISPR-CAS9 system and then , via the Broad institute, licensed the technology to Monsanto.

Greetings forum participants. I’m Matthew Legge and I had the chance to participate in the last online forum as well.

The specific example of malaria control has been raised e.g. [#8427]. Malaria is indeed a horrible health problem for the world community and I would like to believe that no one is callously rejecting synthetic biology solutions due to a lack of understanding about the seriousness of malaria. So I think that listing malaria’s health impacts, while emotionally provocative, is irrelevant to the issue of synthetic biology and biodiversity. In addition, gene drives are being proposed for many other issues aside from just malaria control, so I don’t see the need for us to focus only on the most benevolent of possible uses.

I think the implications for biodiversity may become massive in the near future, should gene drives succeed after either intentional or accidental release (and as has been pointed out, it remains highly unclear if gene drives can succeed). The safety and containment issues mentioned in [#8431] are very significant for protecting biodiversity. It is clear to me that gene drives will be considered and developed to attempt to eradicate entire populations or even entire species unless regulators decide these applications are unacceptable based on uncertainties, contested knowledge, and unknowns.

For me, the acknowledged lack of any international regulatory framework is a huge issue with gene drives possibly impacting on biodiversity. Should any researcher or corporation develop a functional gene drive and release it intentionally or accidentally the resulting impacts on biodiversity (e.g. species decline or loss and all of the resulting impacts on other species and ecosystems) will not respect international borders.

I think that [#8437] raises important points about the question of precision of techniques like CRISPR-Cas9. I believe it is too early to make a general claim of precision (a claim I’ve seen made or implied already, and which I think is often accepted without sufficient evidence) or to know the exact likelihood, scale, or significance of off-target effects. Given the number of unknowns and uncertainties, I have to agree that a precautionary approach at this stage would be to treat genome editing as “highly experimental” due to the paucity of established knowledge about the “off-target effects elsewhere on the genome or epigenome.”

I also agree that comparators for biosafety assessment will become less and less useful as engineered organisms become more and more novel. It will be difficult to even establish levels of risks, which require a number of factors to be reasonably well known. For this reason I am not inclined to agree that we should feel ready to adopt a pro-actionary (rather than precautionary) approach as advocated for in [#8448] because “scientists, clinicians and others have long experience of handling biological agents with potentially harmful effects on human health or the environment...”

I would suggest that these biosafety and international regulatory issues are very important ones that must be fully addressed before the potential benefits of synthetic biology for biodiversity can be realized.

I think this statement is important: “Conservation efforts manage ecosystems to achieve predetermined human values; never to a achieve an outcome of no deleterious effects to all biodiversity.” [#8446]

I find that our actual task is therefore not a technical or scientific one alone. Rather, we are tasked with deciding what level of destruction or alteration of biodiversity is acceptable. That is the first and most important question before we address the technical question of how we can be reasonably certain of not surpassing that acceptable level. In other words, as uncomfortable as it may be, I don’t think the question posed can be answered well without exploring and deciding the core values we seek to uphold with respect to biodiversity and the 3 goals of the CBD.

As I’m on this forum representing a faith community, The Religious Society of Friends (Quakers), I would like to raise that ecological and biodiversity implications of synthetic biology go further than merely can something be done or even can it be done in ways that appear safe or beneficial based on existing (limited) understanding. We would also do well to think about: What does it mean to be in right relationship with other living beings? Is looking for biotech solutions to complex problems through editing life itself the right way to relate to life? In some instances it may be, in others it may be an expression of profound alienation from life, rather than an optimal use of our brilliance and resources. Perhaps adapting to the natural world and stewarding biodiversity through such adaptions of our behaviours, consumption patterns, and so on is in some cases superior to seeking to dominate and alter the natural world through creating synthetic genomes to serve our whims. The CBD can be encouraged to facilitate reflection along these lines, as this is relevant to the topic at hand.

Jim Thomas [ETC, #8437] outlined a number of recent technical or product developments in the general area of synthetic biology.  I think providing this set of cases is a useful stimulus to debate, however I disagree with much of his analysis.  Perhaps it is only fair that, noting “hyperbolic claims” of some synbio boosters, he counters with hyperbolic claims from the other side. Noting (#8439) that these issues have been discussed in depth previously I comment here only on a couple of specific points; some other issues are or will be addressed in other posts.

Gene drives do not attempt to “overcome the principles of natural evolution”.  On the contrary, they attempt to develop and use selfish DNA systems, whether taken directly from an exogenous context (e.g. Wolbachia) or substantially synthetic.  Selfish DNA systems are very widespread in nature and take many forms; much of our genome comprises relics of ancient selfish DNA systems.  See [1] for many examples.

What does the application of machine learning and robotics to synbio mean in relation to the convention?  If multiple changes are made to achieve a given phenotype (e.g. ability of a microbe to synthesis a specified chemical) without direct human involvement in the design, this brings us close to something rather familiar - the product of directed-evolution experiments, or traditional mutation-selection.  These approaches have been safely used for very many years.  Furthermore, in most cases such “microbial factories” are intended only for use in contained fermentation systems and have extremely limited ability to survive outside a cosseted lab setting, not least because of high fitness cost (relative to wild type, outside of specific lab settings) of the many changes made to their metabolic pathways.  #8439 notes a potential exception in GM algae production platforms, which may need “relatively ‘open’…production ponds/facilities”; this simply highlights the much noted need for case-by-case assessment.


1. Burt A, Trivers R. Genes in conflict: The biology of selfish genetic elements. Cambridge: Belknap Press, Harvard University Press; 2006. 610 p.

On “off-target” effects

Several posts have touched on “off-target” effects [see Tony James #8398 for definitions of off-target and non-target], in general considering the possibility of off-target effects as a potentially serious negative issue.  This was perhaps put most strongly by Jim Thomas [#8437]: “Until such a time as it can be shown beyond reasonable doubt that a new ‘editing’ approach is entirely predictable with no off-target effects elsewhere on the genome or epigenome, it would be precautionary to treat genome editing as highly experimental and not ready for commercial or ecological release”

The most readily understood off-target effects, and the most relevant in terms of CRISPR/Cas9 systems, are DNA sequence changes (mutations) induced by cutting of the Cas9 nuclease at a site in the host genome other than that to which it was nominally directed by the guide RNA(s) provided (that being the intended or “target” site).  Other off-target effects are conceivable, e.g. stable epigenetic changes; for simplicity I will here discuss only sequence changes.

Off-target effects of this type are universal in traditional breeding methods, selection experiments, and other (non-CRISPR/Cas9) genetic engineering approaches such as most transgene-integration methods.  Furthermore, even in the absence of any such manipulation, there is a spontaneous mutation rate (this rate varies dramatically between taxa, being extremely high in some viruses, much lower in higher animals, for example). 

One attractive feature of CRISPR/Cas9-based methods is their precision.  Contrary to [#8437], it is not the case that “the hype of ‘precision’ is foundering on the reality of off-target and unexpected changes at the genomic level…”.  Rather, these methods provide a new level of precision and accuracy in changes to the target sequence.  Lack of change in the rest of the genome is an impossible goal, given spontaneous mutation.  The degree to which this background rate is increased by use of CRISPR/Cas9 is not currently clear but some effect should be anticipated.  Importantly, the structure of CRISPR/Cas9 mutations (e.g. SNPs, smaller and larger insertions and deletions) is not different from the normal spontaneous mutagenic range.

How much does a degree of off-target sequence change (mutation) matter?  This varies enormously by application, again highlighting the need for case-by-case consideration.  For human gene therapy, the hazard of oncogenic (cancer-inducing) mutations seems severe, at least to an outsider to the human gene therapy field such as myself, so it would be extremely important to minimise the likelihood of inducing one.  At the other end of the spectrum consider a gene drive system intended to impart a fitness load (for population suppression) into a mosquito population.  There a degree of off-target mutation would arguably increase efficacy by increasing the induced fitness load, though to such a small degree as to be insignificant.  Perhaps more importantly, the level of off-target mutation is not yet clear, and presumably subject also to technical manipulation [#8398], such induced variation is likely insignificant relative to pre-existing natural sequence variation [for the purposes of this discussion I assume dispersed cutting rather than very specific off-target effects, e.g. from a poorly-designed guide RNA that induces efficient cutting at an additional site in the genome other than the target (drive) site].  As a consequence spontaneous mutation and other natural processes, wild populations have significant levels of genetic variation - for example whole-genome sequencing of 765 wild-caught mosquitoes identified over 50 million single nucleotide polymorphisms (SNPs) across the 230 million base genome [1].  Individual mosquitoes carried between 1.7 and 2.7 million variant alleles. 

There may also be useful precedents to consider in this area.  Radiation-based Sterile Insect Technique (SIT) programmes irradiate mass-reared pest insects to sterilise them then release them to mate with the wild pest population.  Radiation induces DNA damage (mutation); if sperm or eggs are sufficiently heavily irradiated then they become non-viable, hence suitable radiation doses can be used for to sterilise insects in this way.  Such SIT programmes have been safely and successfully conducted for more than 50 years, some on very large scales, such as the continental-scale elimination of the New World screwworm from USA, Mexico and Central America as far south as Panama.  Since radiation weakens the insects in a dose-dependent manner, radiation doses are typically selected to give high, but incomplete, sterility, e.g. 96-99% (one exception is that higher doses may be used for preventative releases where the pest is thought absent).  Consequently, many billions of heavily irradiated (and hence heavily mutagenized) but partially fertile pest insects have been released to mate with wild pest insects over many years.  I am not aware of any report of harm to human health or the environment from such introgression of novel mutations, despite the very much greater mutation rate in this context than conceivable from off-target effects of CRISPR/Cas9 or other synbio manipulations.


[1] http://www.biorxiv.org/content/early/2016/12/22/096289

Dear members
#8413- (everything damages environment. So we will damage too) – this may not be the right approach to analyse any technology. This is like the man is already smoking so put him in polluted condition. If something is wrong then we are here to rectify this.

Resurrection of a species is different and preservation of diversity within a species is totally different. Here when we talk about biodiversity we should think about protecting the diversity within the species too. It’s not advisable to instruct a common man that “let everything what nature has donated be destroyed and you please procure the patented goods for your survival”.

thanks

I would like to comment on  the GMO 2.0 Ingredients database posted by Silvia Ribeiro, Latin America Director for ETC Group. I have just had a browse and the database is an excellent resource both for synthetic biology and biotechnology communities and other interested stakeholders -
see http://database.synbiowatch.org

Browsing through the date  there are many petrochemical examples (acrylic acid, acetone butanol  etc) with many companies (and researchers) developing biologically based fermentation methods for producing such compounds with the aim of reducing the dependency on petrochemicals.

Dear participants,

First of all I would like to express my gratitude to the organizers and moderator of this forum. Like others, I also took part in the conversations in 2015.

I am a lecturer (Assistant Professor) in Synthetic Biology at the University of Surrey in the United Kingdom. I concur with many of the views expressed. I must admit that as an active researcher in the field I feel we are, in general, being overcautious with the technology. In my opinion, the benefits of some applications of engineered organisms (including environmental release) can outweigh their potential risks. For example, I see how using synbio to remediate plastic or paracrine hormone pollution can have a positive impact rather than negative in biodiversity. Actually, I feel that if we don’t take action soon there will be very little biodiversity to worry about in less than two generations. I think that eventually the discussion will not be about if we should be taking any risks but, rather, how much risk are we willing to take.

We would obviously need to consider each case individually, but technologies like microbial bioremediation have been around for over 40 years including experiments of controlled release. This means that data is available to evaluate potential impacts and there is the possibility of adopting the same protocols for heavily engineered organisms as well.

I agree with previous comments highlighting the potential of CRIPR-Cas9 as a transforming technology. In this sense I also believe that the beneficial uses of the technology outweight misuses. Society, and I am thinking of countries in which the biotech industry is not too consolidated, would benefit from rapid adoption. The current legislation and patent laws seem to be hindering the progress of existing SMEs and the emergence of new ones in places that in which diversifying economic sector could have a highly positive impact in the GDP.

Hello everyone; I am Jim Louter from Canada.  First of all, thank you Caspar for taking on the task of leading this discussion and for providing guidance to keep it on track (and which you have repeated in post #8442)!  We all need to keep your guidance in mind when replying in this discussion in order to have a logical, progressive and focused session. Like you, I was also a member of the Synthetic Biology AHTEG and participated in the discussions since then; I am also an active regulator (for nearly 25 years) of the products of biotechnology in Canada conducting many environmental risk assessments in that period.  Your suggestion for three points to keep the matter in focus when replying is worth repeating here:
“1) What are the potential negative impacts, including unexpected and significant adverse effects, of the most recent technological developments in synthetic biology on biodiversity and the three objectives of the Convention?

2) What research and cooperation activities are being conducted on the possible benefits and potential adverse effects of organisms, components and products of synthetic biology on biodiversity to fill knowledge gaps and identify how those effects relate to the objectives of the Convention and its Protocols?

3) Are there other recent technological developments that have taken place within the field of synthetic biology that need to be considered in this discussion?”
Regarding the 1st, as Caspar indicated in his opening remarks in post #3865, there have been submissions made regarding these questions including comments from Canada in which we indicated that, to date, there is no evidence of any adverse effect on biodiversity that was identified in a risk assessment that one could attribute to a living organism product of  ‘synthetic biology’.  For the one example I am aware of that involved multiple biochemical pathway manipulations for a microorganism to be used in containment, we concluded there would be no impact on biodiversity.  If we want to speculate about potential negative impacts, I’m not sure where that speculation would end or what utility such speculation would serve. As a risk assessor in Canada, we would be alarmed and would take action where the science told us that an adverse impact of any product of biotechnology, including those that are naturally occurring, was reasonably foreseeable.  In Canada, we are not overly concerned whether or not something is labeled as ‘synthetic biology’ but if it is ‘new’ or it is ‘novel’, that is enough of a trigger for us to begin the risk assessment process. We have a review process that is essentially technology independent. We are however in the process of re-looking at our regulations from the point of view of ensuring that they are up to date – not to redefine the approach (focus on product, and not the process) but, if needed, to refine the information requirements needed to conduct the risk assessment.
Regarding the 2nd, we tried to answer this question with the submission from Canada referenced above but it is difficult to capture all of the research being conducted in one country for the simple reason that ‘synthetic biology’ may not be the correct term used by researcher’s to characterize their activity. Nevertheless, even a simple search would reveal that the Federal Government’s ‘Genomics Research and Development Initiative’ includes many examples of research, some of which could involve synthetic biology and for which the objective is to better understand biodiversity. Canada’s Concordia University has a Centre for Applied Synthetic Biology in which both potential benefits and risks of the technology are being explored.   Both Genome Canada and Ontario Genomics also contain much information on the current state of related research in Canada.
Lastly, regarding your 3rd point, in such a field where technological developments may be fast moving, (and disregarding whether they are labelled ‘synthetic biology’ or not), one would have to look at the research links above to determine whether there was something recent that warranted further attention.  It is in our human nature to innovate. However, there is the danger inherent in focusing only on a few select ‘buzz words’ like synthetic biology – i.e. that a new technology will come along that does not fit the operational definition and we will have to begin this debate again.  Isn’t it better to have a technology independent regime if possible?

Others have made similar comments and I want to support views expressed in posts 8449, 8454, 8448, 8427 and 8410.

Since my hyperlinks have not displayed in this message, I am attaching the Word version with correct hyperlinks embedded.

Thanks once again to Casper for moderating the forum and to the Secretariat for providing this opportunity.

The discussion has advanced quite a bit since yesterday, and I want to offer my gratitude to participants who have shared substantial and informative posts.  I will miss some, but I am particularly supportive of the details and the context in posts #8461, 8454, 8449, 8446, 8439, 8448, 8427 and 8447.

I wanted to especially support some of the points made by Jim Louter in #8461.  One of the challenges of any label is that it seems to imply that anything under the label is materially similar.  Applied to something as broad as synthetic biology it leads to the very mistaken assumption that just by the nature of being synthetic biology something poses a particular risk to biological diversity.  This is not factually correct, and can be easily demonstrated with examples.  A gene edited bacteria in a biofermentor being used in food production has nothing in common with a gene drive introduced into a rat on a small island for the purposes of population suppression with respect to potential adverse effects on biodiversity. 

Although this may seem obvious, one of the manifestations of this fallacy is a tendency to be hyper vigilant with respect to changes in molecular biology.  This is observable in the last twenty years of regulation for biotechnology where huge amounts of molecular characterization data are assembled and assessed with no demonstrable relevance to impacts on the environment or biodiversity.  Just because we can observe and enumerate single nucleotide changes in the billions of base pairs constituting an organisms genome doesn't make these changes relevant or meaningful to biodiversity.  Before a genomic change can harm biodiversity, a number of things also need to occur which can only be determined in the context of the specific organism and its use. So while the potential of any tool (like CRISPR) to cause unintended changes in the genome is important to understand, those changes are not necessarily risky in the context of biodiversity.  And they tend to distract from more worthwhile considerations.

Likewise, I think some of our participants are in danger of falling into the trap of defining any change as an adverse impact.  As has been pointed out, genomes are plastic, ecosystems are fluid not static and the only constant is change.  That doesn't mean that we should not consider potential adverse impacts, but the suggestion that changes to individual organisms and their unique genomes in the environment should be considered an adverse effect is wholly impractical and not in line with the way Parties to the convention make decisions about the environment and biodiversity in the real world.

One of the best tools to improve our understanding of what is, in practice, considered an adverse effect and what is not is to compare the potential impacts of a particular synthetic biology application with other interventions that affect similar elements in the environment.  For example, I don't think it makes sense to agonize over the genetic integrity of pests and disease organisms in order to protect them from genetic interventions if we are just going to turn around and kill them with more conventional means.  So the context of the activity, our management goals, and alternatives are all essential in understanding and contextualizing the potential for adverse effects of any synthetic biology application.

Dear Casper, Secretariat and participants

With respect to the suggestion by our colleagues from the Dutch National Institute for Public Health and the Environment and our moderator [#8442], I disagree that question 1 is better formulated by mentioning “the potential for both positive and negative impacts of the most recent technological developments in synthetic biology as well as of their applications” [#8439].

We are discussing syn bio from a risk assessment perspective. The prevailing instrument already mentioned in this forum is the Cartagena Protocol, particularly Annex III “Risk Assessment”.

The Parties did not include an equivalent annex for ‘benefit analysis’ nor has there been developed any Guidance for how to identify and assess potential benefits.

The better part of a decade occupied another AHTEG on developing Guidance for risk assessment. Nothing of this scale of effort has been asked for by the Parties, nor available for us to refer to in guiding this discussion.

We certainly could fill the many electronic pages of this forum with suggested benefits. Fortunately, there are other venues for these. Assembling a list of benefits may be useful for some, but it is unclear to me how it is useful to fulfill the work objectives requested by the Parties.

Regards
Jack

Dear all—

Respecting Mr. Heinemann’s opinion of the task of this forum [#8463], I would like to provide some excerpts from our terms of reference, as stated in Decision CBD/COP/DEC/XIII/17 adopted by the Conference of the Parties:

1. Building on the previous work of the Online Forum and Ad Hoc Technical Expert Group… as well as information made available through the online forum… the Ad Hoc Technical Expert Group on Synthetic Biology shall…
(a) Review recent technological developments within the field of synthetic biology to assess if the developments could lead to impacts on biodiversity and the three objectives of the Convention, including unexpected and significant impacts…

(c) Further analyse evidence of benefits and adverse effects of organisms, components and products of synthetic biology vis-à-vis the three objectives of the Convention, and gather information on risk management measures, safe use and best practices for safe handling of organisms, components and products of synthetic biology...

As a member of the 2015 AHTEG, I would also like to point out the Boxes on “Potential benefits”  and “Potential adverse effects” included the AHTEG’s report.  It is a good starting place for all of us when considering categories of potential benefits and potential adverse effects.  And by doing so within this online forum, we can be of greatest help to the AHTEG meeting to follow next December.

Regards,
Bob Friedman

Hi! I am Elpidio Peria, from the Philippines, with  the Biodiversity Management Bureau of the Department of Environment and Natural Resources, an agency that oversees the implementation of the CBD in the country.

I would like to highlight # 8392, # 8395 and this one which highlighted the potential reduction of our dependency on petrochemicals, as another, how should we call  on, jump-off point on which to identify what are these potential negative impacts and unexpected and  significant adverse effects.  

When looking at these impacts, #8410 is right that this should be based in reality and #8413 said  we should be looking at specifics.  But have there been sufficient  studies done already on these specific impacts, it seems it is  too early to say and the current exchanges we are having in  malaria  shows how we can further advance  our thinking  on these issues.

My foremost concern at the moment when looking at these potential studies of adverse impacts is there geography, maybe  in the summary, our dear Moderator Casper might be able to summarize who is doing what and where, since  looking for example  at  the literature  cited by #8435 or #8438, these are mostly driven  by countries that are already well-invested  in science infrastructure, which begs the question on whether our moderator's # 2 question may merit enough replies, but even then, these cooperation activities, who drives them, who defines the need for these cooperation activities, or are these potential cooperation activities merely extensions  of the current efforts of protagonists on this issue.

I would also credit #8437 for suggesting a new scheme on which to order our discussion on these technological developments and just singling out #8445's focus on food applications already triggers further questions and perhaps possible concerns that we need to flag down.


Thank you and hope to get back to the exciting and  enriching discussion before this part of our discusison ends.

While Jack Heinemann in #8463 is saying we are discussing synbio from a risk assessment  perspective, I would highlight the importance of looking beyond that framework considering its possible limitations especially when we look particularly at the unexpected and  significant impacts as helpfully pointed out by Bob Friedman in #8464. While the box that Bob suggested is useful, and  we should put  out something like that in this Forum’s  report for the Parties, what we should be looking at  as he pointed out from the COP decision on what we’re doing now, is the evidence, if any, of these benefits.

An example of these unexpected and significant impacts could be what Paul  Freemont  mentioned in #8457, like  “reducing dependency on petrochemicals” , the likes of which we should  foresee when we answer question 1 though there may be things that we cannot adequately foresee and even our information and monitoring of  these LMOs released into the environment as  envisioned by Cartagena may not capture all these unexpected and significant impacts.

What I am saying is that, given the various possible impacts of these most recent developments as helpfully schematized  by Jim Thomas,  which were noted by some of us here as hyperbolic, and indeed we need  to scrutinize which  of those claims will actually hold true, what we may need to eventually consider is whether  the case by case approach of Cartagena may still be a sufficient approach for dealing with these latest developments, especially when some of these new developments may result in  ecosystem-level changes involving multiple organisms,  and we haven’t brought into play the possible implications when these applications enter into the food-stream for humans.

As we are set to finish this coming week our exchanges in question 1, we may have to list down these potential negative impacts but also the unexpected and significant adverse effects and  what  are the studies that are looking at these, and we should point out areas of impacts that are not studied as well as  where are they doing it (that was part of what I was saying in # 8465 when I mentioned “geography”) and put them side-by-side with these latest developments as helpfully re-schematized by Jim Thomas.

Impacts on ABS have been  largely overlooked  by most of us in these  exchange as I suspect this has  not been really extensively  studied as compared to the 2 other objectives of the CBD, but let’s thank Joseph Vogel for keeping at it and hopefully Casper will kindly include  it in his summary as many developing countries Parties to the CBD  and the Nagoya Protocol will be looking for it.

Thank you for the opportunity to be involved in this forum and contribute to the discussion. I have been involved in opportunities over the past few years focussing on policy associated with genetically modified organisms particularly insects.

One of the critical issues in developing proportionate regulations for new synthetic biological (synbio) technologies is to ensure that these are commensurate with the risks. Fundamentally this involves formulating plausible pathways to harm, define the risk (as the hazard multiplied with the exposure to the hazard) and working to mitigate this risk.

A plausible pathway to harm would be expected to have strong risk-based hypothesis. It is also entirely feasible that benefits can be considered with this framework by the appropriate choice of comparators to evaluate the efficacy and biosafety of the synbio technology.

Focussing on insect vectors of disease, it is hard to see how a modified vector would lead to a plausible pathway to harm. An appropriate comparator might  be the levels of the disease under status quo and this would be compared to the novel synbio technology. It is unlikely (as there is no plausible pathway to harm) that as noted in other posts on the forum that these technologies would have an adverse effect on human health (and as such convey a benefit for disease mitigation). This defines an epidemiological paradigm (endpoint) for regulations

Alternatively, for a biodiversity perspective we might define an entomological endpoint to assess the harm to wider biodiversity. Simple epidemiological theory tells us that reducing (and not eradicating) vectors below an entomological threshold will reduce disease transmission and hence affect levels of disease burden, without the full ecological consequences of species loss (irrespective of the invasive status of vector - e.g. Aides). But this still necessitates that a plausible pathway to harm - that the loss of a disease vector will have a greater effect on disease burden/biodiversity than the status-quo. It is hard to envisage this being true (given as noted - the current ways we control vectors).

Developing proportionate, evidence-based regulations predicated on plausible pathways to harm, risk assessment and risk management has to be the way forward.

Prof. Bonsall’s advice in [#8474] coheres with “the economics of uncertainty”, which includes a rigorous literature on psychological biases in perception that was cited in awarding the 2002 Nobel Memorial Prize in Economics. Policies informed by that literature often recommend obligatory insurance due to the phenomenon of cognitive dissonance: confusing a high expected value of a low probability event (the risk)  with its low probability (the hazard) and acting as if the expected value were also low.

As the valuable posts make evident, understanding the multitudinous complexities of biosafety will require specific expertise in many fields. Incentives to engage such expertise and evaluate the relevant complexities could emanate from a governmental requirement to be insured. I hasten to add that such insurance should not have a limit on liability, as any such limit would cynically exploit cognitive dissonance, as happened in the US Price-Anderson Nuclear Industries Indemnity Act of 1957.

Should the Conference of the Parties be informed by “the economics of  uncertainty”, an anomaly would emerge:  “the economics of uncertainty” is conjoined to “the economics of information” under “information, knowledge and uncertainty” in the Journal of Economic Literature (JEL) classification codes. However, the “the economics of information” has been studiously ignored in the CBB and NP (Oduardo-Sierra et al. 2012) . Indeed, no reference to the “economic of information” appears in the heralded ‘The Economics of Ecosystem & Biodiversity’, largely because the object of R&D was (mis)identified as “genetic materials” in Article 2 the CBD.  As the lead TEEB author makes clear at the outset: ‘In the TEEB assessment, we largely follow the definitions of the United Nations 1992 Convention on Biological Diversity’ (de Groot 2010 p.15).

The foundational error reverberates.


de Groot, R. (2010) ‘Integrating the ecological and economic dimensions in biodiversity and ecosystem service valuation’, The Economics of Ecosystems and Biodiversity: The Ecological and Economic Foundations, http://www.teebweb.org/2013-08-30_archive/wp-content/uploads/Study and Reports/Reports/Ecological and Economic Foundations/TEEB Ecological and Economic Foundations report/TEEB Foundations.pdf

Omar Oduardo-Sierra et al. (2012). “Monitoring and Tracking the Economics of Information in the Convention on Biological Diversity: Studied Ignorance (2002-2011). Journal of Politics and Law, http://dx.doi.org/10.5539/jpl.v5n2p29

I found the comment of  [#8474, #8475] insightful for the discussions for the larger society beyond the scientific community (possibly with ramifications for other forums).  It is with relevant scientific article  links.

The issues of "time" (or duration of discussion) is a dimension that plays a key role, in my view.

Dear Members,

I support the views of Prof. Bonsal and Prof. Vogel. In addition to that "genetic material" should also be emphasized along with “biodiversity and ecosystem” many of the genetic material used in R&D are not available in nature. Some of them are developed by continuous efforts of breeders or the farming community.

thanks

Dear Casper, Dear fellow participants,

My name is Margret Engelhard, I am working at The German Federal Agency for Nature Conservation in the area of GMO Regulation and Biosafety and I was a member of the Synthetic Biology AHTEG that met in Montreal in September 2015. 

I would like to comment on the rich discussion on Gene Drives. So far the discussion was focused on vector control for human disease that is at the forefront of this development. In addition I would like to draw the attention to other applications that should be discussed in parallel as their impact on biodiversity might be equally big, including local and global concepts of Gene Drive-technology. Aims for the use of synthetic Gene Drive-Technology (e.g. using homing endonucleases like CRISPR/Cas) include population control in varying different fields of application, e.g. eradication of rodents from islands or pest control in agriculture.

To evaluate their impact on biodiversity some general thoughts might be helpful: The creation of LMOs using synthetic biology differs from the use of LMOs in agriculture (GM-crops) and are predicted to impact biodiversity for a number of reasons:

1) Modification of wildlife becomes both feasible and attractive regarding the multifaceted applications proposed for Gene Drives.

2) Proposed use of Gene Drives on biodiversity issues will inevitably impact biodiversity, either by achieving the desired goal or by unforeseen negative impacts due to problems of the technology, its desired outcome or its application in the wild.

3) Gene Drives moves fields of application of synthetic biology from predominantly contained use to a huge variety of ecosystems (even skipping agroecosystems where GM crops are supposedly to be limited to).

Best regards,
Margret Engelhard

POSTED ON BEHALF OF KENT REDFORD
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Dear Colleagues:

I am Kent Redford with a background in conservation and an interest in the intersection of synthetic biology and conservation practice.
Thank you to all who have contributed - it is a rich discussion.

I would like to enter with a couple of points:

1) when we think about possible outcomes of synbio interventions it is important to compare those possible outcomes to what would happen/is happening in the absence of the intervention. This perspective allows to see that the status of much of biodiversity in all its attributes and components is decreasing steadily - any given intervention may either accelerate or decrease that trend. But preventing interventions will not, in and of itself, make things better.

2) it might be useful to consider potential synbio impacts, both negative and positive, as either direct or indirect. Considering negative impacts, direct negative impacts could be generated by loss of genetic diversity and indirect negative impacts by loss of ecosystems through placement of algal raceways. Considering positive impacts, direct positive impacts could include controlling invasive disease threatening endangered species whereas indirect positive impacts could be through increased food production allowing for less land under agriculture and therefore available for restoration of natural habitat.

Thank you for the opportunity to contribute.

Kent

POSTED ON BEHALF OF RUTH SPENCER
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Dear Colleagues, I am Ruth Spencer from Antigua and Barbuda joining in the discussions from the perspective of the Indigenous peoples and local communities ( IPLC's) and seeing the implications for the ABS/Nagoya Protocol. Our local strains of seeds, plants and other genetic resources have a way of adapting to the impacts of climate change so introductions  and knowledge of  new and diverse species and their impacts  are very important. I am learning from all the discussions and thanks for your sharing but please remember in all  of the discourses that  the ecosystems that we are naturally  blessed with that we are trying to maintain must have priority.

8474--------I fully agree with Pr. Bonsall that developing proportionate, appropriate and evidence-based regulations predicated on plausible pathways to harm, risk assessment and risk management is really the way to be forward.

My name is Mark Tizard, I work with CSIRO, an Australian federal government research agency, and have joined the forum late due to international travel commitments. Thank you to Casper for moderating this forum and to everyone for the enthusiastic contributions thus far.

Topic 1 (Reviewing recent technological developments within the field of synthetic biology to assess if the developments could lead to impacts on biodiversity and the three objectives of the Convention, including unexpected and significant impacts) is our groups focus until 17th July. Clearly the CRISPR/Cas9 gene editing tool and an application of the tool, gene drive, has been a strong focus of post since these are the most significant advance in the field in recent times.   However across the discussion that has been running over the past few days there seems to be a lack of distinction about the three key areas of synthetic biology – at one end this includes gene editing (very minimal change which may mimic natural sequence variation), through “classical” genetic modification (GM) (with clear rearrangements or new introductions of new DNA elements or coding sequences) in the middle and with gene drive (GM with an associated system to push novel DNA based traits into a naïve wild population or gene pool) at the far end. Another important consideration for our deliberations is where the synthetic biology takes place and/or has its use – contained in a laboratory or secure facility (research or industrial production facility), in an open environment but bounded by geography (field bounded GMO crops, or activities on isolated islands or in defined water bodies) or ultimately in a fully wild and unbounded natural environment.  So to evaluate the risk “synthetic biology” present to “the three tenets of the CBD” it is critical to understand for any given case the form of the technology being used (GE, GM, GD) and where it presents any inherent risk (contained, open or broadcast).

It is only natural that much of the discussion has centred on gene drive in a broadcast sense. This is of course the case for which we have the least information. So there is a good deal of research being conducted to fill the knowledge gaps, which is conducted in appropriate physical containment and with geographic containment (in a location where there is no native population of the same species). The potential for risk to biodiversity is only extant after broadcast release and that is only going to happen after extensive risk analysis based on data from laboratory based and other studies. Release can only occur following extensive regulatory approvals.
Post #8486 by Delphine Thizy gives an excellent description of the background to gene drive in relation to the mosquito vector of the malaria parasite. The objective is not to eliminate the mosquito but to reduce its ability to carry the parasite. The answer to post #8479 by Ms Yasin relating to the pathogen (malaria or Dengue or Zika) is that it is not possible to apply synthetic biology to the pathogen because there is no means of disseminating a debilitating trait through the pathogen population (which is dispersed and locked up inside the mosquito population – not even the malaria parasite). 
 
The regulations that should apply would follow the suggestion by Pr Bonsall (post #8474) being proportionate, appropriate and evidence-based and predicated on plausible pathways to harm, with risk management commensurate with the risk assessment. This is in preference to the concept solely of “stringent regulations” (post #8480) unless the risk analysis shows that proportionate and appropriate is equal to stringent.

POSTED ON BEHALF OF Hiroshi Yoshikura
----------------------------------------------------
My name is Hiroshi Yoshikura, once the Codex chair of the Task Force on foods derived from modern biotechnology. Our chair, Dr. Casper Linnestad, asked us three questions, which attracted many interesting comments. If the same questions be asked several years later or even next year, there will be different comments reflecting advancement of the technology involved. In this sense, it may be important for CBD expert group(s) to follow closely the development of technologies related to synthetic biology. Having said, however, despite of new technologies that may arise one after another, the  approach to the risk assessment of the new emerging technologies should remain the same, which is based on Annex III Risk assessment of Cartagena Protocol, particularly paragraphs 5, “risks … should be considered in the context of the risks posed by the non-modified recipients …”, as no living organisms, notably humans(?), are risk-free. For CBD, it may be interesting to consider how this approach, “comparative safety assessment”, could be used for assessment of ever progressing synthetic biology. One approach could be to use as a comparator “conventional counterparts”* that were used or experienced with good or bad consequences now and in the past. 
Sincerely,
Hiroshi Yoshikura
National Institute of Infectious Diseases Japan, Emeritus member

I support Mr. Tizard's proposal:

The regulations that should apply would follow the suggestion by Pr Bonsall (post #8474) being proportionate, appropriate and evidence-based and predicated on plausible pathways to harm, with risk management commensurate with the risk assessment. This is in preference to the concept solely of “stringent regulations” (post #8480) unless the risk analysis shows that proportionate and appropriate is equal to stringent.

Joaquim A. Machado

Dear all,

my name is Christoph Then, I took part in this AHTEG from the beginning. I am representing ENSSER (The European Network of Scientists for Social and Environmental Responsibility, http://www.ensser.org/) in this discussion, my 'homebase' is Testbtioch, a non-profit organisation in Germany http://www.testbiotech.org)

I try to answer to the three questions, being aware that many interesting things were introduced already.

(1) Regarding the question of possible adverse impacts. I would like to illustrate the problem using a recent example published in the context of research on Golden Rice: Crossing the manipulated rice with the Indian variety Swarna led to unexpected gene disruption (http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0169600). The resulting plants showed extensive disturbance in their growth. The new gene constructs interfered with the plant’s own gene for producing growth hormones, and the additional gene constructs were not, as intended, active solely in the kernels, but also in the leaves. This led to a substantial reduction in the content of chlorophyll that is essential for vital functions in the plants. These unintended effects were not detected in previous investigations, and it had been assumed that the genetically engineered plants used in these trials would show genetic stability. In fact, these detrimental genomic effects remained undetected until the transgenic plants were crossed with the variety known as Swarna, which is grown widely in India. If this effect had occurred after large scale cultivation, it is likely that the relevant genetic information would have been transferred to weedy rice known to exchange genetic information with cultivated rice. The problematic gene constructs from the weedy rice could then have been passed into regional rice varieties. This sort of geneflow could have a major negative impact on seed biodiversity.

I think this example is relevant for the discussion of the more recent applications in SynBio: Synthetic biology plans to introduce new technically inserted genetic information and / or technically induced patterns of genetic changes into domestic and wild populations. Organisms, such as those belonging to insect species that are kept in the laboratory, are selected according their potential to survive in the lab. If the new genetic information is passed from these insects into wild populations, the genetic background can be vastly different and lead to effects that were not observed in the lab. Further, they would be exposed to a wide range of environmental stressors that are known to have the potential to trigger unexpected effects in genetically engineered organisms. In many cases, these effects cannot be predicted to the extent that would be necessary to perform sound risk assessment.

Some conclusions: In the light of these uncertainties and unknowns, cut-off criteria have to be defined for risk assessment and regulatory boundaries established to prevent releases that can cause technical genetic information to persist in wild or domestic populations. This is especially relevant for the most recent technological developments, such as gene drives, that are planned to be introduced into mosquitos, pest insects, weeds, rodents and others. There are also other organisms with technically inserted genetic information and / or technically induced pattern of genetic changes that should not be allowed to spread these genetic conditions into domestic and wild populations. These aspects are closely interrelated with potential spontaneous transboundary movements and the protection of agricultural systems, such as organic agriculture, which have to be considered under the objectives of the CBD.

(2) I am involved in a pilot project called GeneTip. http://www.genetip.de/index.php/en/biottip-pilot-study/. If anyone is interested, I would be pleased to provide further background information.

(3) I think several of the issues, such as gene drives and CRISPR have already been mentioned.  In regard to gene drives we have to be aware that possible applications are not restricted to malaria (I wonder if there is some kind of campaign going to propagate this specific application within this online forum?). I also would like to add other techniques, such as Oxitec´s RIDL or the application of miRNA that are meant to interact with natural populations via synthetically derived nucleotides.

Further, the use of gene synthesis to resurrect or create deadly viruses is a hot topic that is currently being discussed in the context of Canadian research on the horsepox virus: http://apps.who.int/iris/bitstream/10665/198357/1/WHO_HSE_PED_2015.1_eng.pdf?ua=1. As already mentioned, this should also be dealt within this group.

In response to Christoph Then (#8495). I am very sorry for the ignorance, but isn't this the ideal case in which when the engineered variety crosses with wild-type does not produce a very fit progeny? It is unlikely that the crosses will take over any population due to their impaired physiology, which eventually will lead to their extinction in the case of accidental crossing.

Very recently I had the chance to attend to conferences on xenobiology (in which natural components of living beings are replaced by artificial ones that can only be found in the lab; e.g. non natural nucleic acids or amino acids), that essentially acts as a measure to prevent unwanted spread of modified organisms (mainly bacteria). Using multiple xenobiological elements is believed to be the ultimate form of biocontainment due to the additive effects of each of the modifications.

Best regards,

Jose (Lecturer in Synthetic Biology, University of Surrey, UK)

Mart Loog, professor of molecular systems biology, Estonian Centre for Synthetic Biology
I agree that synthetic spieces using orthogonal coding and well-defined multiply-backup-ed suicidal containment moduls written in their genome, could be the ultimate and ideal solution. In this case we have to wait until the technology is good enough... before we test any synthetic life in open field.

Chris Then uses “genetic information” six times in [#8495] even though we have been implored to stick to the AHTEG definition which deals with “genetic materials, living organisms and biological systems”.  Not one use of “genetic information” in [#8495] could have been substituted with “genetic materials”, “living organisms” or “biological systems” and retained the intended meaning. The observation is not a criticism of the valuable insights provided by Dr. Then. On the contrary, it is to show once again why we must grapple with correcting the AHTEG definition.

In response to Mr Thomas’s comments re off-target effects of gene editing tools (post #8450), it is important to consider what those off target effects may result in genetically and phenotypically.  The biodiversity that we all seek to defend and protect has come from millennia of mutational events that create large numbers of variant for a majority of genes that collectively instruct the assemble of a given organism in a species and that individual has one composition of those variations of the genes – one set of alleles (the variants of the genes). Natural selection for variations in gene by environment over time (long periods of time) leads to effective congregations of genes (within individuals) that grouped together define species. Without this we would not have the biological diversity we see around us. And it is an on-going process.

What is the point of this? Gene editing (call it CRISPR, that is the current mode but there will likely be other forms in the future, it was TALENs and ZFNs in the recent past) gives scientist unprecedented precision to make directed changes to genomes. Yes precision for the selected target but what of the off target effects? There is conflicting evidence of the frequency with which they occur – it will depend on the organism and the details of how the editing was performed [were you editing this with a laptop, a tablet, a smartphone, pen and paper]. So there may be some unplanned off target edits. Are they important? That question has to be framed in the context of how they might differ from edits caused by natural mutation events that are very frequent in sexual reproduction. Look at a chromosomal spread of gametocytes (the cells that make sperm and ova), look at the number of chiasma that occur – these are the so called “crossings-over”, they are breaks in the chromosomes that are then re-healed after exchange of DNA from maternal to paternal chromosomes. This is the engine-room of genetic variation and reshuffling and it is why sex is so prevalent and so important in the natural world of biodiversity. Breaking and fixing chromosomes is a natural part of this and it is how genetic variations between individual in a population are generated. It is essential to the diversity that is required for the fitness, resilience and survival of a species in a changing environment. 
So we can use whole genome sequencing of a gene edited organism to look for off target changes but distinguishing the few that might come from off target events from those that occur by chance (natural variation) is almost impossible in anything other than an inbred strain of animal or plant.
Chiasma that occur during gamete formation enable gene reshuffling and this is the “genetic modification” that has been used to shape all of our agricultural animals and our companion animals, dogs and cats.  These are “natural” process that have been going on for around 10,000 years.  How do the natural variants of a given gene survive and propagate? First by not being detrimental to the development, physiological processes or behaviours of organism that is carrying them, second by imparting some (albeit) small advantage at some stage in the organism’s life.
If this in an agricultural setting then the risks should be low since the material is controlled.
So the question is if a gene editing system (CRISPR for example) or the introduction of a transgene (GM) has an off-target consequence then what impact does it have on the primary organism (the individual that may be released into the environment)? If it is not deleterious the unexpected change may be retained. If it is retained does it confer an advantage at the expense of others in the population? If so how is this any different to the evolution of any other novel trait based on an allele variant in the population? So does this change significantly alter the role of that organism in the ecosystem in any perceivably different way to any other allele for any other gene in individuals in the population?
Finding and characterising the small changes in the billions of base pairs that make up most plant and animal genomes not sensible way to go in tackling this issue. Probabilities are low that any change like this would have an effect any different to the chances of a similar outcome from a natural variation process.

Responding to Mark Tizard reply (post #8507):  Thank you Mark for your patient and thorough analysis.  I agree completely!

I would like to just add some examples of unintended effects that have recently been documented in LMOs, the same risks could be extrapolated to gene editing techniques, with resulting ecological/health/agronomic effects: 
-Insecticidal Bt Cotton: increased susceptibility of root fungal disease caused by altered levels of sugars and amino acids (Li et al., 2009); reduced levels of Bt toxins during flowering period and   altered chemistry of mature plants reduced toxicity of Bt  toxins to pests (Olsen et al., 2005)
-Herbicide-tolerant NK603 maize: altered composition of nutrients in plant, including 28-fold rise in polyamines – can be toxic (Mesnage., 2016)
- Golden rice: when bred to Indian rice variety, it was found that homozygous plants for the transgene, were dwarfed and stunted due to transgene disruption of gene expression related to photosynthesis and growth hormone production.

Whether or not these disruptions are deemed natural or not fails to take into account that these crops are sold to benefit small farmers in developing countries. Such effects on agronomic performance is only going to exacerbate food production ,especially when such seeds are aggressively marketed over indigenous varieties. This is highlighted by the recent phasing out of GM cotton in Burkina Faso, a nation renowned for its quality cotton. Introducing the Bt transgene into local varieties led to shorter cotton fibres and a decline in profits for the farmers who then pursued compensation for their losses. It is vital that a holistic consideration of the effects of these modifications are taken into account, including socio-economic considerations that might result from unintended effects.

Dear Members
#8510- Always in an organism genes work in a network. When we introduce a new gene, there could be lot of chances of feedback regulation at post transcriptional and translational level within a cytoplasm or organism as a whole. Only case by case (each event is totally different. So we can’t say everything as a success or a failure) analyses will prove its impacts.
Thanks Ms.Delphine (#8486),
#8513 – Ms. Eva – conventional mutation breeding is escorted by selection cycles in natural breeding conditions – (not in open pollinated state). Further, only favorable phenotypes are selected – there is no harm in that. Apart from that, many of the cultivated varieties are mutants and polyploids derived through such procedure.

thanks

Dear Colleagues,
My name is Nikolay Tzvetkov and I work at the Bulgarian Ministry of Environment and Water. I also had the honour to participate at the AHTEG meeting in 2015.
Joining the discussion relatively late allows me to make a relatively short intervention as a lot of information has already been provided. I apologise that I won’t cite many of the previous interventions, but that’s preferable to missing someone.
1) What are the potential negative impacts, including unexpected and significant adverse effects, of the most recent technological developments in synthetic biology on biodiversity and the three objectives of the Convention?
To the best of my knowledge no significant adverse effects on the biodiversity and the three objectives of the Convention related to SynthBio Organisms or SynthBio techniques have been observed so far. In general whether an effect is negative or positive is a matter of values. Most (including myself) will agree that reduction of biological diversity or introductions of invasive alien species in not a good thing. In this case removal of IAS from an ecosystem will be a desirable development. Unless such a removal results in further reduction of biological diversity, a possibility even when the IAS is not keystone specie, but due to secondary and tertiary effects.
Effects on biological diversity can be related to:
1. The biological properties of the organism per se and on its ecological properties in the receiving ecosystem. As the discussion so far seems to show most of SynthBio organisms developed so far are used to produce different chemicals and as such are not very likely to survive in non-controlled environments (some may even carry unnatural components that make survival impossible). In general it seems very unlikely that any SynthBio organisms will have advantages that will allow them to establish in the environment and/or transfer genetic information to other species that will result in significant effects. Exceptions are those organisms that have been engineered in such a way that they should have effects on the environment. Such are the organisms that carry Gene Drives. Although currently Gene Drives don’t seem to as effective as initially expected in changing the genetic structure of populations, we can expect that those technical issues will be solved in near or mid-term future. Such technologies are probably the best candidates for developing tools for environmental risk assessment in international contexts as they have the potential to address really important issues like spread of important diseases, removal of IAS that cause great damage to the biodiversity. At the same time they can be very hard to remove from the environment and they can affect countries way beyond those where they were released. So proper tools should be developed that will allow relatively safe deployment of the technology without sacrificing the benefits.
In general, introduction of any new technology has been and is associated with unforeseen risks. It is a matter of managing those risks to a level that is deemed acceptable by the society. If the stringent criteria and strong precaution proposed by some participants in the forum for products of modern biotechnology are applied to crops or medicinal products developed through traditional means this may result in great inconveniences to put it very mildly.
2. Indirect effects associated with the properties of the organism. Such effects can be increased use of plant protection products, faster exhaustion of soil nutrients, etc. None of those effects seem to be specific for synthetic biology though and it should be considered as done for the traditional products.
3. Effects coming from broader changes in the society due to the introduction of the technology (social and economic consequences). That’s not unique for synthetic biology. Any new technology can and historically has affected negatively certain social groups. That can result in negative impact on the environment due to changes in the social and economic behaviour (there are many such examples). A the same time experience shows that trying to block introduction of new technology on the basis of social and economic concerns is not effective. The much better way forward has been, and probably will be in the future, to allow fair share of benefits of the technology to be redistributed to those negatively affected by it. This seems to allow more people to be better of overall that bans and prohibitions.
Perhaps ironically it was the technologies that produced greatest benefits for the humanity, such as vaccines, medicinal products, fertilisers, plant protection products that have resulted in the biggest damage to the environment. As result of them the people live longer, the population density increases, the quality of life and personal consumption goes up, all that increases the pressure on the environment and damages the biodiversity. But I guess very few people will argue for ban of those technologies on that ground (or any other).
3) Are there other recent technological developments that have taken place within the field of synthetic biology that need to be considered in this discussion?
Here I can hardly add anything more than Boet Glandorf and Jaco Westra (#8439) and Jim Thomas (#8437,) although I don’t share his pessimistic outlook.
Regarding the discussion on Gene editing tools, I would like to point that this is not a synthetic biology development per se, but an enabling technology that has other applications as well. In general I would consider it a mutagenesis tool (it can do other things as well, but the discussion seems to focus on the changes in the genome). Just like any mutagenesis reagent it results in a spectrum of mutations, some (many) of them off target (if possible to talk for on-target mutations with chemical reagents). Gene editing is much more accurate and results in fewer unexpected changes than most other techniques. So we should take this into account, but I see no reason to be much more worried and to require higher level of precaution than with traditional techniques.
Regarding the definition of Synthetic Biology, I am very happy to see that the discussion moved away from it. I am personally not very happy with the current definition and I have advocated in the previous forum one that focus stronger on the creative, engineering, going beyond nature aspect of Synthetic biology. But ultimately I feel from my past experience that those researchers and companies working in a given field care little about names (unless this helps to secure funding from one source or another) and stay focus on discovery and innovation, no matter how it is called.

Dear Participants of the Online Forum on Synthetic Biology thank you for posting so interesting point of views, examples and comments.
I would like to thank also to Casper Linnestad for moderating this topic and the Secretariat for their compilation
My name is Lázaro Regalado MD, Specialist in Microbiology and Member of the AHTEG in September 2015; I’m going to participate in this forum on behalf of the Cuban National Center for Biosafety (Ministry of Science, Technology and Environment). We deal with the issues of regulation, safety and control representing national authority for Biosafety and Biosecurity.
I’m sorry due to my late participation; the reason was the attendance to a National Workshop regarding the design of facilities with biological risk.
Having said that I would like to raise some general issues regarding our first topic
In the Secretariat SYNTHESIS OF VIEWS IN RESPONSE TO PARAGRAPH 10 of
DECISION XIII/17 ON SYNTHETIC BIOLOGY there are different shades of opinions reflecting the complexity to obtain evidence that support benefits or adverse effects.
However in my view we should take into account the preamble of the Convention on Biodiversity:
¨Noting also that where there is a threat of significant reduction or loss of biological diversity, lack of full scientific certainty should not be used as a reason for postponing measures to avoid or minimize such a threat. ¨
In this regard the Cuban Ministry of Science Technology and the Environment does not prohibit, but controls, activities related to the development of new technologies in the biological sphere in general, including synthetic biology if necessary.
To fulfill our task it’s very wise to be guided by the three questions of our moderator meanwhile as it is my first participation I suggest to look at issues such as:

 Unexpected mutations after CRISPR– Cas9 editing in vivo. NATURE METHODS | VOL.14 NO.6 | JUNE 2017 | 547
 Digital-to-Biological Converter for On-Demand Production of Biologics Developed by Synthetic Genomics, Inc.
The first fully automated machine to convert digital code into functional biologics without human intervention creates entirely new avenues for precision medicine
http://www.prnewswire.com/news-releases/digital-to-biological-converter-for-on-demand-production-of-biologics-developed-by-synthetic-genomics-inc-300464948.html
 THE NATIONAL ACADEMIES PRESS. Preparing for Future Products of Biotechnology
http://www.nap.edu/24605
              Committee on Future Biotechnology Products and Opportunities to Enhance
               Capabilities of the Biotechnology Regulatory System
 How Canadian researchers reconstituted an extinct poxvirus for $100,000 using mail-order DNA By Kai KupferschmidtJul. 6, 2017 , http://www.sciencemag.org/news/2017/07/how-canadian-researchers-built-poxvirus-100000-using-mail-order-dna

Respectfully,

Dr Lázaro Regalado

Dear all,
at first I would like to thank Casper Linnestad for accepting the undoubtedly challenging task to moderate this online forum. My name is Swantje Strassheim, I work for the German Federal Office of Consumer Protection and Food Safety in risk assessment of genetically engineered organisms and more specifically on synthetic biology. I have already participated in the last online forum and also followed the discussions at SBSTTA and COP13.
What I think is really difficult about this discussion is the fact that when we talk about synthetic biology, there are always two views: the researchers´ view where synthetic biology is a concept that aims at standardizing and building new artificial genetic circuits, genomes and entities; and the regulators´ view where the interest lies in the question whether organisms produced with synthetic biology can be regulated with the existing tools or not.
Therefore, from my regulator´s point of view, I would like to answer the questions Casper has posed:

1) What are the potential negative impacts, including unexpected and significant adverse effects, of the most recent technological developments in synthetic biology on biodiversity and the three objectives of the Convention?
Many participants in this forum have stated genome editing techniques as CRISPR/Cas, TALEN, zinc finger nucleases etc. as recent technological developments of synthetic biology. However, I would like to differentiate a bit more. All of the genome editing techniques, but especially CRISPR/Cas, can be used for a variety of genetic modification. If a double or single strand break is induced by CRISPR/Cas, which is then repaired by endogenous enzymes, there will only be very small deletions or insertions (indels). Whether this leads to a GMO or not is actually extensively discussed and I think we have to wait for the final verdict on this issue. If organisms with these indels are considered GMO, they do not pose any new risk to biodiversity that could not be assessed with the existing GMO regulations, which in case of the CBD would be the Cartagena Protocol. The same applies to insertion or deletion of a whole gene, which is considered to produce a GMO that can also be assessed with the existing GMO framework and has no new risks to biodiversity.
Another technique cited a lot in this forum are gene drives. From a regulatory point of view these are also GMO and can be assessed through the existing protocols. I agree nevertheless, that due to the nature of gene drives more precaution in risk assessment is necessary. The German Central Committee on Biological Safety (CCBS) has issued a position statement on the use of recombinant gene drive systems in containment (http://www.zkbs-online.de/ZKBS/SharedDocs/Downloads/02_Allgemeine_Stellungnahmen_englisch/01_general_subjects/Recombinant%20gene%20drive%20systems%20(2016).pdf?__blob=publicationFile&v=3 ). In its statement the CCBS notes that for risk assessment of a recombinant gene drive system it is relevant, whether the gene drive can get in contact with interfertile organisms of a wild population in the event of accidental escape, if the wild interfertile population contains the specific target sequence, if the inserted nuclease is expressed constitutively or needs a specific inductor/effector and if free and random mating in the population is possible (many domesticated species, e.g. livestock, are bred by selective mating). The CCBS therefore concludes that the “extent and rate of spread of a gene drive system in interfertile, naturally occurring populations depends on a number of factors that need to be identified and assessed on a case-by-case basis”. Depending on the identified factors, a given gene drive could by assigned to BSL-1, -2, 3 or -4 and specific safety measures adapted to the gene drive and its target organism can be issued.
Furthermore, with all the exciting news about the problems gene drives could solve, we have to keep in mind that gene drives might not work the way they are supposed to. The spread of gene drives may easily be stopped by natural species varieties and resistances to the gene drive probably occur frequently (see Paper by Unckless et al. 2017, http://www.genetics.org/content/205/2/827).

2) What research and cooperation activities are being conducted on the possible benefits and potential adverse effects of organisms, components and products of synthetic biology on biodiversity to fill knowledge gaps and identify how those effects relate to the objectives of the Convention and its Protocols?
The German CCBS is been conducting a monitoring on synthetic biology since 2009 and has released its first monitoring report on synthetic biology in 2012. The next report is expected for the end of 2017 and will, just as the first report, come to the conclusion that all organisms created with the help of synthetic biology so far are GMO and can be risk assessed with the existing regulations and guidance. As an example, the CCBS has recently assigned the minimal organism Mycoplasma mycoides JCVI-syn3.0 to risk group 2, due to the currently scarce knowledge about the pathogenic potential of these bacteria. It is however possible that this minimal organism is a lot less pathogenic than the wild type and will not survive in the environment. When these assumptions will be verified a reclassification in risk group 1 can be effectuated.

To answer question 3), from my point of view, there are currently no other technological developments that need to be considered in this discussion. In line with this I agree with Andrew Robert [#8410] and Ethan Bier [#8417] that we do need well-defined examples as to where current risk assessment frameworks are not applicable to synthetic biology before we start discussing any new approach.

Ms Strassheim wrote [#8533] that “In its statement the CCBS notes that for risk assessment of a recombinant gene drive system it is relevant, whether the gene drive can get in contact with interfertile organisms of a wild population in the event of accidental escape…Depending on the identified factors, a given gene drive could by assigned to BSL-1, -2, 3 or -4…”  This seems to me entirely sensible.  However it may be worth considering the possibility of some indirect interaction with Access and Benefit Sharing. If containment issues for some synbio organisms may be more problematic where there are endemic populations of the (unmodified) species, this may mean that work on various species, e.g. pest species but also potentially beneficials, may be more easily or even more appropriately conducted away from the area in which they are present in the environment.  Remote researchers may have a comparative advantage in having lower containment requirements (if the CCBS approach were universally adopted).  Regions with a problem potentially addressable by such methods might prefer the initial research to be conducted elsewhere.  Such corollaries may not always fit well with concepts of local R&D on local genetic material/resources.

Dear participants of the synthetic biology on-line forum,

My name is Ana Atanassova and I would like to thank the Secretariat and the moderator of the forum for the opportunity to participate in this lively discussion forum.

A  number of people have submitted comments about “off-target effects” related to the application of CRISPR/Cas9 and similar genome editing tools. Two recent publications were quoted as evidence of such “off-target effects”. [Unexpected mutations after CRISPR–Cas9 editing in vivo, Schaefer K.A. et al. (2017) Nature Methods, 14(6): 547); CRISPR/Cas9 targeting events cause complex deletions and insertions at 17 sites in the mouse genome, Shin H.Y. et al., (2017), Nature Communications 8:15464].

It is broadly recognized that changes to the sequence of the DNA of an organism, or mutations, occur spontaneously or as a result of  human intervention, and that  induced changes have to be seen in the context of the natural variation and evolution of the genome. Furthermore, not all changes in the DNA have an effect on the functioning or characteristics of an organism. While we debate “off-target effects”,  we should remember that “off-target mutation” is not equal to “off-target effect” and not imply that all mutations lead to negative effects in all cases. I support the comments made in posts #8454 and #8507 that highlight that “off-target effects” are not in any way unique to applications of site specific nucleases such as CRISPR/Cas9 and would like to share a reference to an excellent review on the subject: Schnell et al., (2015). A comparative analysis of insertional effects in genetically engineered plants: considerations for pre-market assessments. Transgenic Res. 24(1):1-17.

Going back to the publications mentioned in the beginning of this post. Schaefer et al.  was widely discussed in the scientific community as not having a proper experimental design to support the conclusions that the authors made about a link between CRISPR/Cas9 and a large number of non-target mutations observed. Critical examination of the experimental design leads to the conclusion that a likely explanation for the authors’ observations is that the detected polymorphisms (off-target mutations) are not CRISPR/Cas9 related but rather evidence of genetic polymorphisms inherited from parents of the two genetically related experimental animals. The links below provide more detailed analysis of the results:
• The experimental design and data interpretation in “Unexpected mutations after CRISPR–Cas9 editing in vivo” by Schaefer et al. are insufficient to support the conclusions drawn by the  authors http://arep.med.harvard.edu/pdf/Schaefer_Opinion_2Jun2017.pdf?from=timeline&isappinstalled=0
• “Unexpected mutations after CRISPR-Cas9 editing in vivo” are most likely pre-existing sequence variants and not nuclease-induced mutations http://www.biorxiv.org/content/early/2017/07/05/159707
• CRISPR Gene Editing Controversy: Does It Really Cause Unexpected Mutations? https://www.forbes.com/sites/stevensalzberg/2017/07/10/crispr-gene-editing-problems-does-it-really-cause-unexpected-mutations/2/#2adfbfdb3ed9

Irrespective of the design weakness of the experiment,  the authors can be commended for concluding  that “researchers should carefully assay their specific gRNA and Cas9 for off-target mutations, before  the CRISPR platform can be used without risk, especially in the clinical setting”. I stress their conclusion as it differs from the way it was broadly reported in the popular press and in this forum as to imply “off-target effects” due to CRISPR/Cas 9, or genome editing in general.  In reality this study, amongst many others, is an example of how researchers are addressing scientific questions to advance the understanding of the technology towards future safe use in human therapies and other applications. Similarly, the article by Shin H.Y. et al., (2017) makes a contribution to the scientific understanding of how best to develop CRISPR/Cas9 for in vivo applications and is by no means a warning about the limitations of the technology.

Therefore, information from basic research should be recognized as contributing towards the understanding and development of a more predictable and safer technology but should not be used to project hypothetical risks for potential future products. Product risks should be addressed on a case-by-case basis as has been the situation for the current commercial products of LMOs.

If i may contest the latest comment on natural modifications.
I agree there is natural modifications that take place in genomes, in the form of transposable elements and horizontal gene transfer etc, indeed it is these natural genetic modification processes that are utilised for artificial genetic engineering. However, natural mechanisms have evolved over millennia to include tight regulatory checks, for example the epigenetic regulation of transposable elements that largely silence TEs, and looking at latest research, some TEs become activated in times of stress or during cell differentiation for example, thus suggestions have arisen that they are a kind of response to generate variation when needed (though this appears to be early days in our understanding). Artificial genetic modification is designed to override such regulation and as such cannot be directly compared.

I urge those further interested in the differences between natural and artificial genetic modification to read the following paper: http://www.mdpi.com/1099-4300/15/11/4748

Finally, with regards to Schaefer's methodology in the recent CRISPR off-target effect paper, it is, in my opinion, a distraction to focus on this paper's limitations, as it is only representing a continuation of what the research has told us thus far. Off-target effects are to be expected, and unintended effects at the target site also, based on unpredictable gene repair mechanisms. It is not contested by CRISPR developers such as DuPont and Caribou sciences, or anyone else in the field, as revealed by the many, many papers working on CRISPR design improvements, development of techniques to detect off-target effects and regular reports of off-target effects.

I would like to comment on the single author commentary paper that was cited for the discussion on natural versus genetic engineering by Ms. Eva Sirinathsinghji. The manuscript presents little scientific  evidence or actual scientific data  to support the authors hypothesis/theory  that all non natural genetic changes are harmful arguing that organisms have a 'fluid genome' and needs genetic changes to survive termed by the the author " an exquisitely precise molecular dance of life ". More importantly perhaps  the manuscript has not been peer-reviewed.

I would warn colleagues to read this paper with great caution and scepticism as it is not founded on current scientific understanding of genetics, fitness and adaptation nor on the detailed molecular mechanisms around genome integrity and stability. Whilst I appreciate the need to consider a variety of opinions, i  much prefer rationale viewpoints based on peer-reviewed scientific evidence.

on my strawberry anecdote (its Wimbledon tennis  in London at the moment and we eat lots of strawberries while watching the tennis), no one has guaranteed me that the strawberries i eat do not have multiple genetic changes that have resulted in large juicy sweet fruit and that if these were released into the wild, all wild strawberries could be adversely affected though gene transfer and cross-overs resulting in woodland full of very large and highly desirable strawberries .

Dear Dr. Paul,
Please read this. Already replied your question.

Topic 1-conservation of biological diversity - natural variations contribute to diversity [#8514]
thanks

Dear Casper, dear members of the forum,

Thank you for organising this forum.

This contribution follows the French High Council for Biotechnology’s major work published on 7 June 2017 on the use of GM mosquitoes for vector control, available online at:
HCB Scientific committee report:
http://www.hautconseildesbiotechnologies.fr/sites/www.hautconseildesbiotechnologies.fr/files/file_fields/2017/06/06/aviscshcbmoustiques170607.pdf
HCB Economic, ethic and social committee report:
http://www.hautconseildesbiotechnologies.fr/sites/www.hautconseildesbiotechnologies.fr/files/file_fields/2017/06/06/hcbceesmoustiquesrecommandation2juin2017.pdf
the translation of which will be shortly available.

With respect to Casper’s three guiding questions, this touches upon questions 1) and 2), as a research and cooperation activity conducted on the possible benefits and potential adverse effects of SB on biodiversity, as well as other questions, as explained below.

We would like to highlight two key points:
- the importance of specific-case (evidence-based or scenario-based) analyses rather than broad statements,
- the informative value of comparative analysis.

Our work aimed at characterising and identifying interests and limits of applying mosquito control methods using GM mosquitoes, a work that we carried out in a comparative manner with other control methods. We considered:
- different conventional methods (specific examples of chemical, biological and environmental methods)
- different emerging methods, such as sterile insect technique using irradiated mosquitoes (classical SIT), techniques using Wolbachia-infected mosquitoes for population reduction (Incompatible Insect Technique) or population modification (Pathogen Interference propagation), Oxitec RIDL approach (considering the example of Ae. aegypti OX513A line, developed by Phuc et al., 2007) as well as CRISPR-Cas9 gene-drive based methods for population reduction (considering the specific working example developed by Hammond et al., 2016) or population modification (considering the specific working example developed by Gantz et al., 2015).

Questions addressed in a comparative manner included:
1) ability to achieve specific goals : population reduction (or suppression) versus population modification (or alteration), at different scales and degrees (ex: local elimination vs species eradication)
2) efficacy, durability and sustainability (including questions of different types of resistance evolution and genetic drift, inherent limitations of efficacy in space and time, constraints related to initial field conditions such as population density, etc.)
3) technical constraints (including requirements for mosquito mass rearing conditions, separation of males and females, etc.)
4) risks to the environment and public health (including broad questions such as the degree of specificity of the different techniques and associated consequences on public health and non target organisms, potential risks associated with resistance evolution and genetic drift, and more specific questions for different sets of techniques -- for gene drive, the question of off-target mutations and their case-specific consequences, etc.).

Kind regards,

Catherine Golstein and Jean-Christophe Pagès
Scientific Officer and Scientific Committee President
High Council for Biotechnology (HCB)
Paris, France

One may disregard the single author review because it potentially isn't peer-reviewed, but plentiful peer-reviewed data exists and was covered in the review, to show that predicted risks regarding GMOs have largely materialised over the last 20 years, highlighting the continued need to take precautionary measures as we enter into the latest in GM and synthetic biology developments.

Disruption of gene expression resulting in agronomic/ecological/health concerns have been documented, the generation of novel nucleotides, alterations in compositional profiles (inc. hugely increased levels of potentially toxic molecules - see Mesnage et al Scientific reports 6, 37855 (2016)), instability of transgenes, pleiotropic effects, combinatorial effects of multiple transgenes, and so on.

As previously mentioned, such unintended effects have had real impacts on farmers and economies. The introduction of Bt transgenes into high quality cotton in Burkina Faso is the latest example. Introgression of the transgene into local varieties was performed, resulting in the vast majority of cotton in country becoming GMO in just a few years. Last year they had to phase it out because it had reduced the cotton fibre length and thus harmed the quality and subsequent profits for small-scale farmers who then sought compensation from Monsanto for the failures.

Hello everyone. My name is Jenna Shinen and I am the Life Sciences Specialist in the Biodiversity Division, in the Office of Conservation and Water, at the U.S. Department of State.  I serve as an adviser to the U.S. government on international policy considerations related to synthetic biology and work with the U.S. National Focal Point to the CBD.  I am pleased to join the discussion on the forum, and thank the moderators for their work and the thoughtful comments of the other participants in the forum.

The United States understands synthetic biology as it is discussed in the research and development community to encapsulate a continuum of biological engineering tools and techniques leading to progressively advanced biotechnology products. Research in the field of biological engineering improves our understanding of biological systems and contributes to efforts addressing food security, environmental, energy, and health challenges.  Indeed, over forty years of research, education, and product development using recombinant DNA techniques have led to clear benefits relevant to the Convention’s objectives, and these benefits will continue to emerge with continued application of biological engineering tools and techniques.

For example, recombinant human insulin was first licensed in 1980 and is now used worldwide to treat diabetes in humans.  Medical research with transgenic mice and other organisms produced through biological engineering has enabled the elucidation of diseases and therapies for humans and animals.  Peer-reviewed, independent studies have demonstrated that genetic engineering has improved crop production methods by reducing soil erosion, decreasing fuel and chemical pesticide use, increasing disease- and pest-resistance within plants, increasing on-farm insect biodiversity, raising crop product quality, and improving farm productivity and farmer income.

There is a broad range of applications for synthetic biology technologies, including public health/medicine, agriculture/livestock, industrial uses, species conservation, environmental remediation, and invasive species control, to name a few.  There are inherent differences among those uses (e.g., contained use vs. environmental release) as well as different final products. 

This wide array of applications also suggests the need for flexibility and adaptability to address the present set of innovations as well as successive generations of technology.  Reference has already been made to the U.S. National Academy of Sciences report "Gene Drives on the Horizon," as well as a subsequent report on "Preparing for Future Products of Biotechnology."  These are the types of horizon scanning exercises that we will continue to commission, to ensure U.S. domestic and foreign policies keep pace with technological growth and evolution. These reports also provide timely information from multiple sectors with extensive expertise in genetic engineering fields and can be used to inform domestic and international discussions.  Technology development is not static, and our social and regulatory systems will have to adapt to that pace of change.

In fact, new developments in biological engineering technologies are expected to accelerate the rate at which scientists can develop applications of biotechnology to address medical, biomanufacturing, environmental, and agricultural challenges.  These technologies are also revolutionizing biological research, advancing our understanding of living organisms and systems, and are vital to powering the global economy.  At the same time, application of these technologies also brings associated health, safety and security concerns – including the possibility of accidental harm and intentional misapplication. Governments, academia, and private sectors should must collaborate to review governance and oversight mechanisms and address risks associated with applications of genome editing and synthesis technologies in ways that allow for continued innovation and preserve the benefits these technologies can provide.

The United States encourages independent and cooperative scientific research, development, and capacity building in fields relevant to biotechnology and biological engineering, both domestically and with partners around the world. In addition to the extensive research of the private sector, a number of U.S. government departments and agencies fund research in the area of biological engineering, including the Departments of Agriculture, Defense, Health and Human Services, and Energy as well as the National Science Foundation; National Institute of Standards and Technology; and, the National Aeronautics and Space Administration.  Some of the research focuses on filling the gaps in fundamental understandings of biological systems as well as technology development to speed the application of biological engineering and enable commercialization of research.  There are also specific programs in areas associated with stability and evolution of genetically engineered organisms, including mechanisms of containment and biosafety to reduce the likelihood of adverse effects as well as specific programs to examine the relationship between environmental pressures, ecology and evolution.

The National Science Foundation (NSF), in partnership with the Woodrow Wilson International Center for Scholars and the Center for Nanotechnology and Society at the University of Arizona, developed a roadmap for progress in evaluating potential environmental risks associated with synthetic biology and assessing public perception and societal risks and benefits of biological engineering.  Efforts at the NSF-funded Synthetic Biology Engineering and Research Center at University of California Berkeley (SynBERC) addressed environmental risk and societal concerns, the Engineering Biology Research Consortium (the NSF funded successor to SynBERC) continues those activities.  An NSF-wide working group on synthetic biology that includes representatives from the biological sciences, physical sciences, engineering, and the social and behavioral sciences provides a mechanism for coordinating the agency’s efforts in the area of synthetic biology and biological engineering.  Finally, NSF has partnerships with a number of international entities including the United Kingdom’s Biotechnology and Biological Sciences Research Council and the European Commission to fund research in the area of biological engineering and synthetic biology.  In many of these joint research programs, consideration of the responsible conduct of research (including ecological and societal impact) is a review criterion.  There are discussions about increasing such international activities, which could increase research capacity and training in partner nations.

I look forward to a continued, focused discussion on this topic.

- Jenna

Dear forum participants,

My name is Felicity Keiper and I work in agricultural biotechnology research, LMO risk assessment and regulatory policy. I was a member of the first AHTEG on Synthetic Biology that met in 2015. Thank you for the opportunity to comment on this topic.

While the aim of this discussion is to address the topic and guiding questions provided by our moderator, I would like to add my observations on the operational definition that was the outcome of the work of the AHTEG. As noted by others early in this discussion, the operational definition of synthetic biology has been deliberated at length – by an online discussion of this open-ended forum and the AHTEG in 2015, and by the parties to the CBD at SBSTTA-21 and COP13 in 2016. The outcome is a compromise (as noted by our moderator #8377), and not all may be happy with it. This situation reflects the great difficulty of the task, given that in this forum the term “synthetic biology” is being used as an umbrella term to encompass all biotechnological developments post-Cartagena Protocol (e.g. genome editing, gene drives), and in some cases, relabel technologies that were established at that time. This raises the question of why we are even trying to do this, when the CBD already provides a broad definition of “biotechnology” that would encompass the heterogeneous collection of “recent developments” that have been proposed in the present discussion.

“Biotechnology” means any technological application that uses biological systems, living organisms, or derivatives thereof, to make or modify products or processes for specific use (CBD Art 2)

I agree with the many participants (#8410, 8417, 8427, 8435, 8447, 8462, 8465, 8487, 8491, 8492, 8533, 8546) that have emphasized the need to consider specific applications in their specific context on a case-by-case basis. A technology is not itself inherently risky; risks are determined on the basis of the novel characteristics of the resulting organism, its intended use and receiving environment, in comparison to other (non-modified) organisms, on a case-by-case basis (consistent with Annex III of the Cartagena Protocol). I would also add (related to guiding question 3) that this discussion needs to be framed by realistic and foreseeable applications, taking into account current technical capability (as pointed out by #8449), and realistic time frames – is the application merely a research proposal or has it been proven with foreseeable environmental release. A research proposal (i.e. an idea) presented as a “recent development” with speculation of it potential impacts cannot be used to justify the imposition of “stringent” regulatory measures in the name of precaution. I agree with the participants that have emphasized that any regulation should be evidence-based and proportionate to risk (#8452, 8474, 8478, 8491, 8492, 8494) – this is also consistent with the risk assessment principles of the Cartagena Protocol.

In response to guiding question 1, I agree with #8439 that it is repeating work already undertaken by the Open-ended online forum and previous AHTEG in 2015. Rather than speculating on “potential” impacts, the discussion needs to be based on evidence if it is to inform and progress discussions to identify information/knowledge gaps, and fulfill the necessary criteria to justify ongoing work on synthetic biology under the CBD. The information presented in the current series of online discussions is intended to support the work of the AHTEG, which includes providing recommendations to SBSTTA to assist completion of their assessment of synthetic biology as a “new and emerging issue” relevant to the CBD (Term of Reference (e), CBD/COP/DEC/XIII/17 Annex). This assessment requires evidence in the form of credible sources of information, preferably from peer-reviewed publications (UNEP/CBD/COP/DEC/IX/29 paragraphs 11 and 12).

A review of peer-reviewed scientific evidence is presented in the recent information submission of the Global Industry Coalition (GIC). This submission is attached to this post as unfortunately the literature cited was not included the summary (synthesis) of the submissions provided by the Secretariat. For that submission, the Executive Secretary requested “evidence of benefits and adverse effects”, and the GIC submission focuses on evidence for impacts of LM crops as these are the LMOs with which there is the most extensive experience in environmental release (the literature on benefits of biotechnology in agriculture is publicly accessible at: http://biotechbenefits.croplife.org/). What is evident in this discussion is that many of the potential negative impacts listed for synthetic biology today are a repeat of those asserted when LM crops were in development. However, LM crops have been in cultivation throughout the world for more than twenty years without credible evidence of negative impacts on biological diversity arising from use of biotechnology.

As a final point, this discussion greatly benefits from the sharing of real-world experience by the regulators in this forum (see e.g. #8439, 8461, 8533). This is particularly useful for those countries with less experience in conducting risk assessments for LMOs that seek guidance on this. There is more than forty years of accumulated knowledge and experience gained from conducting risk assessments for contained and environmental uses of LMOs. As stated by #8493, existing approaches to risk assessment based on Annex III of the Cartagena Protocol remain applicable regardless of biotechnological developments. This is supported by the findings of the Open-ended online forum on risk assessment under the Cartagena Protocol who considered synthetic biology risk assessment in 2016 (see: http://bch.cbd.int/onlineconferences/2014_2016period.shtml). In that discussion, regulators experienced in LMO risk assessment could not identify or foresee specific examples of organisms that presented biosafety risks that could not be managed using established regulatory approaches for LMOs. It is also evident in that discussion, as well as the recent information submissions, that regulators do not even use the term “synthetic biology” (see also #8461), with LMOs assessed according to existing biotech regulatory frameworks.

We wish to thank the Secretariat for its synthesis of the responses to Notification 25 to inform the online discussions, and Mr Casper Linnestad, our patient moderator who is trying to keep the discussion focused on the three guiding questions.

New Zealand provides its comments within the context of our legislative framework for the regulation of genetically modified organisms (the Hazardous Substances and New Organisms Act 1996). Potential environmental impacts, both positive and negative (including impacts on biological diversity), of any GMO in New Zealand are assessed on a case-by-case basis.

We are of the view that it is essential to evaluate the benefits of any LMO/GMO created through recent technological developments against any potential risks, using robust and effective risk/benefit assessment procedures, as is required in our national legislation. We therefore support the sharing of real life case-studies including both benefits and risks through the CBD’s Biosafety Clearing-House and through this online forum. We acknowledge the real life examples provided and the wide range of research that is being conducted, such as mentioned in posts #8483, #8437, and #8439 (without necessarily agreeing with all the posts’ conclusions).

In terms of responses to new technologies and assessment of impacts, the range of uses described in the posts highlight that any risk assessment framework needs to reflect the actual use of the modified organism and that its use be considered within the context of a particular country / environment. These comments have also been made by for example #8410, #8461, #8462, #8492, #8530 and #8533.

It is not clear from many of the examples provided in the posts how these differ from LMOs/GMOs as they were considered when the Cartagena Protocol was first drafted. Although such posts are within the scope of the very broad description of synthetic biology in Decision XIII/17, we suggest that perhaps a more useful discussion could focus on areas where the synthetic biology technology stands to make some step changes (engineering/standardised parts/automation/etc).

Mariska Wouters
Ministry for the Environment

Dear participants of this forum, I would like to express my sincere gratitude to all of you for being both very active and cooperative. Let me also remind you that at this stage, there are only a few more days left in this first discussion. If there are any final comments you would like to make, please do so soon!

Beyond doubt, you have provided the CBD with a lot of insights. The views you have shared on topic 1 (“Review recent technological developments within the field of synthetic biology to assess if the developments could lead to impacts on biodiversity and the three objectives of the Convention, including unexpected and significant impacts”) to me clearly demonstrates that synthetic biology is something that the CBD must address properly in the years to come. For one thing, the AHTEG will have a lot to discuss at its next meeting!

The closing of this discussion will be done by the Secretariat on Monday morning. At a later stage, summaries from the discussions will be provided by the Secretariat with the aid from the moderators.

It has been a pleasure to moderate this first topic of the discussion. You have been guiding each other very constructively, making this an easy task for me. I have learned a lot. I am also very happy that we do not have to reach consensus or agreed language at this stage….

Sincerely and with gratitude,
Casper

Dear all,

My name is Deborah Scott, and I am a social scientist at the University of Edinburgh. I research the governance of emerging science and technology and related decision-making processes, and I’ve been following synthetic biology at the CBD since 2010. I’d like to connect back to a few points raised in previous posts:

1) In #8496, Jose Jiminez referred to xenobiology as “believed to be the ultimate form of biocontainment.” Last week I attended a workshop organized by Vitor Pinheiro on “Xenobiology: biocontainment, biosafety, and biosecurity” (possibly you were there, too, Jose?). I want to commend the xenobiology community for engaging with these issues at such an early point in the development of their field. For the purposes of this forum, it’s worth noting that xenobiology research is very much focused on conducting basic scientific research at present. Since at least 2009, academic papers have raised the possible use of xenobiology as a mechanism for biocontainment (see Marlier 2009; Schmidt & de Lorenzo 2012; Acevedo-Rocha & Budisa 2016; Kubyshkin & Budisa 2017). Nonetheless, as I understand the current state of science, these have been entirely theoretical discussions. Experimental research in xenobiology has not produced a “biocontained organism.” My sense from the workshop and the literature is that containment is an area some xenobiologists are enthusiastic about, but none would claim to be anywhere close to delivering.  An area for the Convention and its Protocols to keep an eye on and keep abreast of, but not currently a technique for containment.

2) I’d also like to flag cell-free systems, identified in #8439 by Jaco Westra and Boet Glandorf as an area for which risk assessments need further scrutiny. I’ve noticed increasing discussion within scientific communities about the benefits of cell-free systems as a way to avoid regulatory ‘hurdles.’ For example, the report from a February 2017 UK BBSRC-funded workshop in South Africa, “Bakubung Workshop Report: Capacity Building for the Bioeconomy in Africa” refers to cell-free and transient expression systems “as easy to implement, relatively free of biosafety evaluation requirements and cheap to deploy….These new technologies avoid complications, delays and regulatory uncertainties associated with uncontained use of GMOs, eliminate requirements for cold chain setups for transport and storage, and provide new options for high level collaboration on education, training and interdisciplinary research between UK and African scientists in low-resource environments…These novel non-GM approaches offer new prospects for (i) low cost diagnostics and environmental sensors, (ii) programmable cell-free expression systems, (iii) vaccine production for rapid responses to emerging viral threats, (iv) biomining and bioproduction, (v) new breeding techniques in plants based on genome editing using CRISPR/Cas9, (vii) new opportunities for training and education in UK and Africa, and (viii) an opportunity to engage societies concerned about GM technology.”

This excerpt gives a sense of the range of possible uses for cell-free systems, as well as the explicit expectation that such uses will not fall under the same biosafety standards as LMOs/GMOs. The AHTEG may want to consider whether this is the case, and, if so, possible implications.

Thanks so much for your moderation, Casper, and thanks to all for the interesting and lively discussion!

I strongly support the final comments of Ms. Atanassova. Moratorium has been always  a useless procedure, if  we make a rational overview of Technology Coevolution, mainly when applied to Biology.

All the best,

Joaquim A. Machado

Dear Participants,

My name is Bernd Giese and I work at the Institute of Safety and Risk Sciences (ISR) at the University of Natural Resources and Life Sciences in Vienna.

In this forum uncertainties have been mentioned several times with regard to risks of new technological developments in synthetic biology (#8451, 8474, 8475, 8495). The precautionary principle demands that "lack of scientific certainty“ shall not prevent a party from taking preventive measures.
In this regard it would be worth to think about a classification of the events that have been discussed according to their level of uncertainty. Such a differentiation is not meant to distinguish between harmful and less harmful events.
But to get a clear picture of what may be avoidable and what will remain it could be helpful to distinguish in our discussion between
a) events that will exist further on (inherently coupled to a certain technology),
b) adverse effects that may become avoidable or at least minimized by further improvements,
c) unknown events that cannot be excluded but their investigation is much to complex and expensive

For this purpose one could consider the (meanwhile) well known scheme of knowns and unknowns:
1) knowns (e.g., extended land use for biomass production and its well known negative impacts)
2) known unknowns (e.g., effects of certain transgenic products (pesticides) on particular species of plants or animals)
3) unknown unknowns (e.g., hazardous (long term) effects on ecosystems due to the eradication of an entire species by a gene drive)

Based on this classification one can either
- decide to have a moratorium until we have found ways to tackle the known unkowns (case 2 above) e.g., by avoiding unintended consequences or at least have appropriate measures to stop a detrimental process or reverse its negative outcomes OR
- decide to live with OR
- not to live with a potential risk and substitute the respective technology by alternative approaches.

The need for cut-off criteria was already mentioned (#8495). And the recently quoted cell-free systems (#8556) seem to be one of the options to reduce the unknowns.

Best regards,
Bernd Giese

Thank you all for the interesting discussions!

1) Post #8483 points to two clear ways whereby the objectives of the Convention are impacted by digital sequence information on genetic resources, i.e. novel pathways for invasive alien species (IAS), and novel invasive alien species (Objective 1: Conservation) and use of genetic resources without benefit-sharing (Objective 3: Fair and equitable sharing of benefits). While the establishment of the AHTEG on Digital Sequence Information on Genetic Resources should go some way in clarifying the issues, discussions under this process (Online Forum and AHTEG on Synthetic Biology) must not lose sight of the issue either, given the role that synthetic biology techniques play.

Many developing country Parties at COP 13 in Cancun proposed to treat digital sequence information as equivalent to physical biodiversity samples for the purposes of benefit sharing. There may be a future need to relook at or supplement the operational definition, and this forum should certainly note the concerns and analyses raised around the issue of digital sequence information and its potential negative impacts on the objectives of the Convention, while mindful of any potential gaps that might arise from the two processes dealing with the issue and ensuring that they are dealt with in a coordinated manner.

2) Much has been said about gene drives as a new development in the synthetic biology field and the knowledge gaps around the science. This is certainly an area of active research and the concerns raised about the potential to irreversibly alter wild populations, species and ecosystems in the absence of clear international governance arrangements and risk assessment procedures are very valid.

The Africa Group at COP 13 in Cancun laid out the issues well, points that were also taken up by civil society: The application of the precautionary approach in the creation of gene drives in the laboratory until gene drive-specific regulations for biocontainment are developed and implemented, the application of the precautionary approach in the consideration of release of gene drives until thorough risk assessments are performed, including of ecosystem and socioeconomic risks, as well as the need to inform and obtain consent from other governments whose biodiversity could be affected by any proposed gene drive before approval of its release. This is by no means a blanket call for a moratorium, but a call to take pause and not proceed until the appropriate safeguards are in place, and to ensure precaution.

The issue of contained use needs to be addressed to ensure that the regulations in place are fit for purpose and can deal with the specific risks that gene drive organisms may pose. This has been highlighted thoroughly by several posts. Guidance on risk assessment aspects of gene drives, identified previously as a specific area of focus by the AHTEG on Risk Assessment and Risk Management of LMOs convened under the Cartagena Protocol on Biosafety, also need to be developed further, while taking into account socio-economic, cultural and ethical considerations. This work could be initiated by the AHTEG on Synthetic Biology given the agreement that risk assessment methodologies may need to be updated and adapted for current and future developments and applications of synthetic biology. In any case, it should certainly be discussed in this forum.

kind regards
Lim Li Ching
Third World Network

I am Benson Mburu Kinyagia from National Commission for Science Technology and Innovation,  Kenya and a member of Syntetic Biology AHTEG.

I agree with[#8449]  that regulatory and governance structures need to be established to the emerging technologies and applications and with #8474, #8491, and #8494 that there is need to develop them in a proportionate, appropriate and evidence-based regulatory framework that is predicated on plausible pathways to harm, risk assessment and risk management. Both positive and negative impacts are relevant to the discussions as they are to to the objectives of the CBD.

Dear Collegues :

I have read with interest the many comments on this review of new biotech. I have been encouraged by the sensible practical approach of many and alarmed by the dogmatic and unscientific comments of others.  I would just like to add that I consider the current  risk assessment procedures used in most developed countries for considering LMOs and their products are generally appropriate for assessing new breeding and genetic techniques. Whether they are proportionate and necessary is a separate issue.  In some cases, such as RNAi, the data requirements for risk assessment may be different since the gene product is an interfering RNA and so data is required on its target, nontarget and off target effects in all potential recieving biota and environments. In relation to gene drive,  worst case scenarios may be necessary to simulate local or regional extinction effects on ecosystems and their functions.   In relation to REAL synthetic biology (i.e. the de novo production of organisms from new arrangements of synthetic and natural DNA) the risk assessment is more complex. Traditionally RA is comparative i.e. the LMO is compared with its nearest non-modified form. In the case of Real SynBio there is no non-modified or parental form. Therefore the risk assessment has no baseline from which to measure additional/changed  effects. This is not really a problem with inert products such as foods since there are analytical techniques for assessing food composition and quality.  However for novel persistent reproductive organisms the RA becomes more complex as there is no baseline info on their ecological role and activities.  They would have to be compared with surrogate species considered to be similar and be subjected to a range of  studies within contained biospheres in order to attempt to predict their impacts when released . Several commentators have already commented on this and provided some guidance ( Epstein and Vermiere 2016) . As indicated in the EC report on SynBio it may be necessary to include genetic confinement or kill mechanisms in some novel organisms where it is difficult to predict outcomes from releases.
Finally I repeat  points made by others: 1. New biotech per se has no impact. It is a series of genetic techniques and so it is the products that require assessment. 2 The impact of LMOS both harmful and beneficial will depend largely on how they are deployed and their management. Therefore this apect is important to assess and manage at local and regional levels, as LMOs and their management will inevitably impact (agro)biodiversity.
Jeremy Sweet : Independant consultant and risk assessor, JT Environmental Consultants, Cambridge, UK .

Dear participants,

First I´d like to say thank you for all the interesting contributions and for the moderator´s work in such a challenging task.

Regarding the three questions presented by the moderator: 

In my opinion the relationship between Synbio and biological diversity was intensively discussed in the cycle 2014 - 2016 in the on line forum (http://bch.cbd.int/synbio/open-ended/discussion_2014-2016.shtml#topic1; http://bch.cbd.int/synbio/open-ended/discussion_2014-2016.shtml#topic4), in countries submission (http://bch.cbd.int/synbio/submissions/2017-2018.shtml) and in the AHTEG (Doc UNEP/CBD/SYNBIO/AHTEG/2015/1/3). In this case it will be more productive to concentrate the discussions in this and the next weeks only in the analysis of recent technological developments with some good examples already presented by many participants in this on line forum to assess if those developments present a new challenge for the risk assessment and regulations in place in many countries.

Synthetic Biology is a tool, as many other innovative tools, and it should be used to address some of the challenges facing society in a near future. In Brazil, all organisms obtained by synthetic biology are considered to be GMOs and we consider the current GMO risk assessment framework therefore applicable also for these ‘synbio organisms´, on a case-by-case basis, without any evidence of potential adverse effects in the biodiversity until this moment. Synbio applications are mostly related with bio manufacturing under contained use, a condition that greatly limits any potential biodiversity adverse effect.   

It is also important to mention that although I consider that current risk-analysis tools are robust enough to cover ´synthetic organisms´ at this moment, it´s necessary to monitor he continuous advance of biotec products and, if necessary, implement changes in the regulatory systems. The countries will have to be prepared to meet the three objectives of the Convention while fostering beneficial innovation. We should use all the experience and familiarity we have with two decades of RA of transgenic products to update, accordingly, the reality of the regulatory framework of each country.

Thank you.

Best regards,
Luciana P. Ambrozevicius

Ministry of Agriculture / Brasil

In response to Deborah Scott (#8556),

Yes, I took part in the xenobiology workshop at UCL as well. I agree that little is know about the biocontainment properties of non-natural components of cells and I felt that, in general, more is required, specially in terms of experimental results, to assess the chances of survival of organisms containing these elements. Given the low fitness of lab-adapted organisms such as modified bacteria (also a topic addressed in that meeting), I would expect an high level of containment while using xenobiological functions (low rate of survival) of organisms in the wild. However, this is just speculation and we would need to produce more data. Same applies to 'orthogonal' components of cells, although in this case, evolutionary paths leading to taking advantage of the orthogonal modules and incorporating them to the physiology of the engineered organisms are likely to emerge, most likely with higher frequency that proper xenobiology components.

In any case, I don't think the current lack of knowledge should preclude advancing in the scientific endeavours dedicated at developing these technologies. On the contrary, as I discussed in a previous post, I feel that we are systematically neglecting potential positive impacts at a time in which we really need to provide society with solutions.

Best,

Jose

Dear Participants,

Thank you to Casper Linnestad for moderating the discussions and all participants who bring positive contributions to this forum. My name is Marcelo Freitas and I have been involved in the discussions about Synthetic Bilogy since he first appeared in CBD.


I would like to support and thanks the comments of Jaco Westra and Boet Glandor [#8439], Thomas Nickson [#8446] and Paul Freemont  [#8449] to bring clearing and scientific facts about theses issues.

Hello my name is Hilary Sutcliffe, I run a not for profit in the UK called SocietyInside which explores how to align innovation better with societal needs and concerns.  I participate in the UK Synthetic Biology Governance Sub Group and the advisory board of the SynbioChem centre.

We work across technologies and believe that there are lessons which often don’t get passed from one to the other as most science & policy works in silos.  Observations from our early work in nanotechnology seem to have some relevance here, as the themes of the discussions here are remarkably similar.  There are three main areas I would like to draw attention to:

1 ‘Ologies’, like nanotec & synthetic biologies are ‘brands’.

They are invariably created by academia to attract funding and investment, as well as being held up by governments keen to show they’re supporting the “next big thing”.  The creation of the “ology” of nanotech led to discussions of risk that were driven by the brand of nanotechnology, and not the science behind how new materials actually behave or the potential hazards and risks they present in reality.

A campaigner at the NGO Greenpeace talking about his work in nano governance observed:   “I wish we’d spent less time worrying about the ‘ology’ and more trying to figure out what we really had to worry about and what we didn’t. We probably lost ten years because of this.”  

Looking at this forum I think synbio is in danger of doing just the same.  Both are important to clarify and I applaud the organisers for trying to get to the bottom of this.  Good luck with that Casper!

2 Hype has consequences

Our ability to seriously engage with the question of ‘what are we really worried about and what are we not worried about’ is further clouded by the hype associated with the brand.  Hype, both about perceived benefits and the perceived risks, means much time is spent worrying about risks and ethics of speculative applications which are far away from being realised, if ever.   Some feel that “speculative harms are treated as fear-mongering while speculative benefits are allowed to run wild”, both are unhelpful.

Trying to drill down to a case by case, application by application based approach to benefit and risk seems to me to be preferable to an ‘ology’ based approach, though of course definitions of certain types are essential for governance.   Work in nano based on ‘plausible pathways to harm’, as discussed here, resulted in quite different conclusions to those about brand nano & size related effects.  

Of the many damaging repercussions of hype on all ‘sides’ of the debate one strikes me as particularly important here - regulators have to start early to consider legislation around the risks and hazards of a new technology. The only place they can start is with what scientists and businesses say they will deliver. Those developing regulation in the life sciences have explained to me that, with hindsight, this for them has been significantly problematic, resulting in regulations which are sometimes not fit for purpose.

3    Following the consequences of the hype comes the attempts at normalisation.  ‘Nano is just chemistry, I don’t know what all the fuss is about’, is echoed here in relation to synbio, which adds to the distrust.  There have been criticisms and the usual, I feel rather disingenuous, comments wondering why people think that products produced via natural breeding and other genetic modification techniques are considered OK and those by synbio not.  On an application by application basis some may be different and some may not, patronising language doesn’t help!   The juxtoposition of the much vaunted ‘precision’ of these techniques, with the reality of the current state of understanding, off target effects and potentially unresolvable uncertainties, contributes to distrust.  A focus on drilling down to what is known and unknown as a previous poster has outlined in detail is essential.

But in the end, creating new organisms that don’t exist in nature, and potentially changing eco systems, is special.  It isn’t just about science, it intersects with values, reasonable concerns about scientific hubris & money taking priority over people and the environment.  It isn’t surprising that some comments are ‘unscientific’ because there are other concerns here than just the science of one technology.  In addition, gauging impacts on ecosystems may, according to the beliefs of scientists, be beyond science’s current or near future capability.  Where does this leave us?

I find myself much inspired by a comment from the UK by Baroness Onora O’Neil, who says “…the slightly plaintive question ‘How can we restore trust?‘ is on everyone’s lips. The answer is pretty obvious. First: be trustworthy.  Second: provide others with good evidence that you are trustworthy.”  

Much of the rhetoric and hype about synbio isn’t focused on building trustworthiness and therefore it is not surprising if trust is not automatically bestowed on its applications. Congratulations and good luck to the organisers for initiating this forum to help do that through multi-stakeholder debate.

Many thanks Mr. Luke Alphey for elucidating the non-targeting issues and the functioning of new gene-editing techniques such as CRIPR-Cas. These clarifications are essential for those who do not work directly with the subject.

I totally agree with Mr. Jose Jimenez's comments. Synthetic biology needs to be considered as an important tool for solving major problems, such as:

Public Health through the control or alteration of organisms transmitting infectious agents, such as arboviruses (viral encephalitis, Dengue, Fever Yellow, Mayaro, chikungunya, Zica, Meningitis), Chagas disease, Lyme disease; Control or alteration of infectious agents, such as Schistosomosse; Control or alteration of natural reservoirs of infectious agents, such as rodents and bats;

Conservation through the control or modification of organisms transmitting infectious agents or threatening the survival of other species; Elimination of invasive species or threatening a particular ecosystem and / or biodiversity; Change in threatened or endangered organisms;

Agriculture through the control or modification of organisms that cause disease or threaten plantations; Elimination of weeds or other undesirable crops in plantations; Development of plants and animals resistant to biotic and abiotic stresses; Development of plants and animals for agriculture with higher productivity; Development of plants and animals adapted to climate change; Environmental Sensing; Agents for Biocontrol;

Basic research through the modification of model organisms for studies of diseases, syndromes, signaling pathways and mechanisms of action of compounds / drugs; and

Biosynthesis through the production of fine and high value-added metabolites such as, therapeutic, pro-biotics, antibiotics, enzymes, antibodies; Production of structural materials, such as fibers, biomaterials.

Thanks for your comments Mr. Jim Loter and I totally agree. The submissions of information on Synthetic Biology made by Brazil are well aligned with this view.

Dear participants,

So far, technical issues related to new developments and tools in synthetic biology have been addressed in the forum, and how these developments can solve many problems, and in turn have adverse effects on biodiversity and the environment. In this sense, I would like to discuss aspects of interest that are related to the questions raised in topic 1 and is the fact that the advances and developments derived from synthetic biology impact the scope of the three objectives of the biological diversity convention, This reason I consider the following approaches:

1. How access to genetic resources and derivatives is addressed when genetic and chemical information for the development of components and products of synthetic biology is obtained through bioinformatics.

2. In the case of using genetic and chemical information found in databases of public use to develop components and products of synthetic biology and that this establish access to genetic resources and their derivatives as the provisions and object is enforced Of the CBD.

3. How is the distribution of the benefits derived from access to genetic resources and derivatives derived from the information found in the bioinformatic means of such genetic resources and derivatives, when developing components and products of synthetic biology?

In reference to the closing remarks of the Moderator [#8555], an “agreed language”  could emerge with a different consensus: one on the persuasive power of logic. The rules violated for crafting definitions in the AHTEG definition for synthetic biology [#8393] and the fallacies in its defense [#8444] are compounded when neologisms are aggregated to address the flaws. A good example is “digital sequence information on genetic resources,” referenced in [#8559] and elsewhere in this forum. “Digital” is only one medium of transmission of natural information for R&D, albeit the most predominant. “Digital sequence information on genetic resources”, therefore, does not capture the less deployed media such as print, film recordings, sound-analog recordings and, more fundamentally, gas, liquid and light for smell, sound, taste, touch and sight. Will the AHTEG someday have to add on the medium of print? film recordings? sound analog? etc? At some point soon, fatigue will set in.

Each of the other words in “digital sequence information on genetic resources” can be similarly unpacked, especially the awkward use of the preposition “on”. Wisely, the Secretariat has called for views to be submitted about “digital sequence information on genetic resources” with a deadline of 8 September 2017 (http://www.cbd.int/doc/notifications/2017/ntf-2017-049-abs-en.pdf). What will happen if those views suggest an alternative to “digital sequence information on genetic resources” with negative implications for the AHTEG definition of synthetic biology?

Reductionism is “the virtually unchallenged linchpin of the natural sciences” (Wilson, 1994). Where is Ockham’s Razor (aka The Law of Parsimony) in the discussion about language?

Wilson, Edward O. (1994) Naturalist. Washington, DC: Island Press

Dear participants, moderator, dear all

My name is Juan C. Menéndez de San Pedro López, I’m Microbiologist, MSc in Biosafety. I’m from Cuba, Ministry of Science, Technology and Environment, Former Director of the Cuban National Center for Biosafety (CSB). I’m going to participate in this forum on behalf of the CSB. We deal with the issues of regulation, safety and control representing national authority for Biosafety and Biosecurity.

This is a late participation due to I was with my fellow citizen in a National Workshop as he said, both as professors.

In my first participation I’m going to talk about 2 of the question raised by our moderator, although some general comments about Synbio at the beginning, most of them already expressed by other colleagues:

1. Synbio is an emerging science extremely complex taking into account the specialties and areas of knowledge like in life sciences and technical sciences, that’s why   the diversity of approaches and criteria’s.  With the current knowledge about Synbio and its regulation is difficult to predict with a high certainty the potential positives and negatives impact that’s why many criteria are elaborated on the basis of hypotheses.
2. The biosafety objective is regulate activities that can pose a risk for health (human, animal or vegetal) and the environment in general based on the process of risk assessment, so we need research in this regard and the development of tools. It is precisely these results that can bring us closer to a probable impact and the way to manage it
3. To conclude the general comments I would like to highlight that the absent of evidence about negative impacts originated by Synbio does not meant their absence.

1) What are the potential negative impacts, including unexpected and significant adverse effects, of the most recent technological developments in synthetic biology on biodiversity and the three objectives of the Convention?

As the question suggests, these may be some of the potential negative effects already mentioned by several colleagues, hopefully they will not be generalized:

1. Worsening of the structure and fertility of soils, caused by the development of biofuels.
2. The possibility of invasiveness and unintended effects of xenobiology.
3. Resurrection of species to the detriment of in situ conservation.
4. Dissemination of unwanted traits (invasiveness, alteration of the food chain, toxic effects on soil microorganisms, insects, animals and plants, introduction of diseases) due to the transfer of genetic material.
5. The generation of pollutants through new production methods (bioplastics, extraction of fossil fuels).
6. Radical or harmful environmental changes (destruction of habitat) by the release (intentional or accidental) of new organisms resulting from Symbio.
7. Allocation to conservation and displacement projects of small farmers (by the substitution of natural products with synthetic products).
8. Gene-editing techniques (CRISPR / Cas9) may lead to unexpected and unpredictable changes in the genomic sequence and thus in the phenotype of an organism and its descendants.
9. Use of genetic resources without benefit-sharing.

3) Are there other recent technological developments that have taken place within the field of synthetic biology that need to be considered in this discussion?

I think the examples provided by Ms. Boet Glandorf, Netherlands [# 8439] and Mr. Edward Hammond, Third World Network [# 8483]. In relation to the latter in terms of:

1. The techniques or tools that allow the resurrection of endangered species, especially of pathogens. (See: Kupferschmidt K 2017. How Canadian researchers reconstituted an extinct poxvirus for $ 100,000 using mail-order DNA, Science July 6, DOI: 10.1126 / science.aan7069)
2. Machines designed to facilitate the synthesis of genomes from digital sequence information are rapidly advancing. The so-called "digital-to-biological converter", (Boles KS et al., 2017. Digital-to-biological converter for on-demand production of biologics, Nature Biotechnology, May.ie: 10.1038 / nbt.3859)

I thank the opportunity to participate in this forum,

Best Regards,
Juan Carlos Menéndez

Dear collegues,
My name is Marina Bogdanova and I represent the National Co-ordination Biosafety Centre (NCBC) of Belarus.
I want to express sincere thanks for the possibility to participate in this Forum. I found a lot of useful and interesting ideas and information here in order to improve my own understanding of the problem.
If to talk about the discussion occurs, I want to support the views of Mr. Jim Louter from Canada (#8461) that we shouldn’t concerned whether or not something is labeled as ‘synthetic biology’ and begin the risk assessment process of all novel products.
Mr. Mr. Joaquim A. Machado wrote (#8476) that “propositions of "stringent regulatory restrictions" are a moving backwards with no practical effects”. But after all it is a question not of "stringent regulatory restrictions", and about reasonable regulation and exploration both potential benefits and risks of the technology.
Also, the idea that the moratorium on gene drive research would ensure that the ‘missing knowledge’ is never obtained, given by Mr. Mark Benedict (#8528), seems appropriate.
As to the main question of this Topic, the negative impacts of using synthetic biology organisms/components/products was already described by many and I’ll join to those of the participants, who underlined the negative consequences of using sterility genes and resistance genes markers in edible crops; that any regulation should be evidence-based and proportionate to risk; and that existing approaches to risk assessment remain applicable regardless of biotechnological developments.

Thanks

Dear Ms. Sutcliff, thanks for your comments.

As a quantitative geneticist I can't see hype about Synbio, unless when propagated by areas outside the core of Science. If this is the case, during all my very important times with CBD, Cartagena and Nagoya, I have seen lots of hype about Nature Conservancy, highly negative impacts of GMOs etc.
Sincerely, instead of hype about Synbio, what I see is the irreversible Coevolution vector moving ahead. Instead of hype, I see Science.

Wish you all the best,

Joaquim

Dear Colleagues:

To inform and provide context for the discussion on Question 2 "What research and cooperation activities are being conducted on ... synthetic biology", I share a just published co-authored paper on "Tracking the emergence of synthetic biology." Following explanation of the search method, this research highlights the leading locations and disciplinary categories for synthetic biology research. We highlight a relatively concentrated set of research sponsors in funding the growth and trajectories of synthetic biology.  A top group of 20 research sponsors is acknowledged in 70.6% of the more than 3,300 funded synthetic biology articles published between 2009 and 2015. Eight funders among the top 20 global sponsors of synthetic biology research are located in North America, with six each in Europe and Asia. All are publicly funded. Arguably, the policies and priorities of these research sponsors will have a significant influence on the directions taken by the synthetic biology field.

In addition to the attachment, the paper is available as open (free) access at https://link.springer.com/article/10.1007/s11192-017-2452-5

Philip Shapira

Professor of Innovation Management and Policy, and Director, Manchester Institute of Innovation Research, Alliance Manchester Business School, University of Manchester, M13 9PL, UK

Co-Investigator and Lead for Responsible Research and Innovation, Manchester Synthetic Biology Research Centre for Fine and Speciality Chemicals, Manchester Institute for Biotechnology, University of Manchester, M1 7DN, UK.

Dear all,

With respect  Question 2, I would like share to you  this Webpage:

https://www.iso.org/committee/4514241/x/catalogue/p/0/u/1/w/0/d/0

The technical labour in ISO will be relevant in this topic, including the technical precedent.

Best regards,

Dear Casper, dear all

This has been a most interesting discussion and it’s difficult to focus on any single issue within such a spectrum.

So let me for a moment return to Casper’s original questions. Regarding question (3) – which other technological developments have taken place within the field that need to be considered in our discussion – I find the following developments crucial to consider:
- Multiplexing (targeting multiple sites within a genome at once): eg. with CRISPR/Cas9 or PTG/Cas9 (polycistronic tRNA-gRNA) or use of other nucleases.
- Rapid identification and/or development of a multitude of nucleases (eg Cpf1), nickases and fusion proteins for genome editing (eg dCas9 or nCas9 linked to activation-induced cytidine deaminase (AID) or methylases)
- DNA-free genome editing
- Next generation sequencing tools
- Combining (new) genetic engineering techniques and tools
- Gene drives: global gene drives as well as local gene drives (as an application of the techniques and tools, including those mentioned above)
I also read with great interest the recent developments listed by Jim Thomas in [#8437], which will need to be considered in the context given in this forum.

Multiplexing:
Many tools are being developed to enable the simultaneous targeting/altering of multiple sites within a genome in one go/at once, often with just one construct/vector. Recent examples of development or application:
Ordon et al. (2017) are basing their work on a “synthetic biology ‘starter kit’”
“Development of a streamlined toolkit for genome editing in dicotyledonous plants: The modular cloning principle and toolbox recently provided to the plant community as a synthetic biology ‘starter kit’ are at the basis of Dicot Genome Editing (pDGE) vectors developed here (Weber et al., 2011a; Engler et al., 2014). Different types of vectors were generated for streamlined assembly of RGN-encoding constructs.” [RGN= RNA-guided nucleases].  The kit developed by Ordon et al. is designed for quick assembly of up to 8 sgRNAs (single-guide RNAs) at a time.
REFERENCE: Ordon et al. (2017) Generation of chromosomal deletions in dicotyledonous plants employing a user-friendly genome editing toolkit. Plant J. 89, 155-168   http://onlinelibrary.wiley.com/doi/10.1111/tpj.13319/full

Minkenberg et al. (2017) make use of the PTG-system in their multiplexing research on rice, a system enabling, as above, simultaneously targeting between two and eight genomic sites in one go. They explain: “The polycistronic tRNA-gRNA (PTG) gene system is a new strategy to provide stable and efficient expression of multiple gRNAs from a single transcript (Xie et al., 2015). This synthetic gene consists of tandemly arrayed tRNAgRNA sequences driven by a PolIII promoter (Figure 1). Every repeat is less than 180 bp long, in comparison with the at least 500 bp needed for an individual cassette.”
REFERENCE: Minkenberg, B., Xie, K. B., and Yang, Y. N. (2017) Discovery of rice essential genes by characterizing a CRISPR-edited mutation of closely related rice MAP kinase genes. Plant J. 89, 636-648  http://onlinelibrary.wiley.com/doi/10.1111/tpj.13399/full



Nucleases, nickases and fusion-proteins enzymes:
The constant development of new and altered enzymatic (fusion)proteins that can be directed to specific genomic sites with CRISPR is allowing on one hand for a much wider array of target sites (due to different PAM sites or spacer length) as well as for not just creating blunt or overhanging DNA double strand breaks (eg Cas9 or Cpf1), but also for cutting simply one strand (nickase), or enzymatically changing a nucleotide (eg deaminase) or causing epigenetic alteration and gene silencing by methylation of nucleotides (methylases).  These have multiple implications for the speed, quality and quantity of (potential) applications and associated risks.


Other:
I do not want to go into further detail at this stage about the new developments (other than having mentioned them), as I find the important question with regard to the CBD and its underpinning three objectives is (1), trying to ascertain the potential negative impacts, including unexpected and significant adverse effects, of the most recent technological developments.

There has been a substantial debate in this forum already, so I will limit my comments briefly to:
- off-target effects
- process induced genome alterations (mutations)

and will add three observations:
- Plants, animals and micro-organisms are increasingly viewed as or reduced to genetic resources for the purpose of “mining”, such as allele mining, and in this resorting to the full set of ‘omics’, including transcriptome-based analysis (reviewed for vegetable crops in Cardi et al. (2017) Genetic Transformation and Genomic Resources for Next-Generation Precise Genome Engineering in Vegetable Crops. Front. Plant Sci. 8.).  There are implications for the three objectives, with indications of a shift away from the first two objectives towards specific aspects of the third. We may need to look at drivers and (perverse) incentives that may undermine the focus and efforts on conservation of biodiversity (incl. agrobiodiversity) and related ecosystems. At a time of severe biodiversity loss, what are the implications of a “mining approach” ?  
- The speed of developments and of the applications of new techniques, either alone or in combination with each other, may become too great and too pressing as to allow sufficient time for reflection, for experience and observation, or -critically-  for evidence to emerge. Risk understanding and risk assessment require time. We need to ensure this time is given or can be taken.
- It appears to me that assumption or unwarranted broad extrapolations are increasingly taken as facts (eg re precision, predictability, reliability, safety or even necessity), which is counter to our tasks at hand. This is also crucial in relation to gene drives (whether global or local).



Concerning off-target effects:
There has been an intense discussion here starting with Barbara Livoreil’s comment early on, and, if I am not mistaken, there is a clear acknowledgement that off-target effects are a reality and a problem that practitioners of genome editing are trying to overcome. Whilst there might be disagreements on degrees, or which experimental procedures would give the most reliable results, or which mice perhaps should be or have been chosen (which by the way is a recurring theme in the often heated debates around LMOs and the accompanying pesticides used), there are clear indications that we have an issue at hand that needs way more research, time, observation and understanding before conclusions can be drawn.  In the meantime, given the precautionary principle, we need to act on the basis that off-target effects do occur regularly, even at places that were not predicted, ie in places that are not captured by the current algorithms.

Indeed that was an aspect that I found greatly helpful in the recent Schaefer paper (see end of para). I am fully aware that aspects of this paper have been criticised, but it is asking some of the right questions. For example it questions our reliance on or belief in algorithms (“Algorithms generate likely off-target sites for a given gRNA, but these algorithms may miss mutations.”), as well as pointing out that indeed we work with assumptions, such as “the widely accepted assumption that CRISPR causes mostly indels at regions homologous to the sgRNA”.  This assumption has led to many statements that there are “no off-target effects”, and the abstracts of many papers will assert this absence of off-target mutations, but when actually reading such papers it will become evident that only a limited number of sites have been checked by sequencing, at times only three.  Whole genome sequencing (in fact deep whole genome sequencing) is the only basis for scientific claims of no off-target effects.
For reference: Schaefer KA et al. Unexpected mutations after CRISPR–Cas9 editing in vivo. Nature Methods (2017) May 30;14(6):547-548. doi: 10.1038/nmeth.4293

Given the history of science and of technical development we would be ill advised to make assertions about the “precision and predictability” of current genome editing techniques, incl. CRISPR/Cas9. Claimed certainties of safety and of no-harm have left trails of suffering and serious harm in their wake, as so well documented in the report “Late lessons from early warnings: the precautionary principle 1896-2000”. https://www.eea.europa.eu/publications/environmental_issue_report_2001_22

If helpful I am happy to provide further papers and links illustrating and acknowledging the off-target effects of site directed nucleases (SDNs), esp. CRISPR/Cas9, or the lack of whole genome sequencing, though I find the links and references put forward by Eva Sirinathsinghji and also by Jim Thomas illustrate sufficiently the current reality of off-target effects. In any case, it is the consequences of such off-target effects that we need to address here, as helpfully pointed out by some.

I apologise for the delay of my contribution. I had wanted to comment on other individual points raised, and also to remind us all of the reality of process induced genome-wide mutations arising from processes of genetic modification and transformation (see Wilson et al. Transformation-induced mutations in transgenic plants: analysis and biosafety implications. Biotechnology and Genetic Engineering Reviews 23 (2006): 209–237 http://econexus.info/publication/transformation-induced-mutations-transgenic-plants).  However, I ran out of time.

There is a wealth of different comments & references made in this online forum, which I hope we all can build on.

With very best wishes to you all,
Ricarda

My name is Gerd Winter, a law professor with some experience in the regulation of biotechnology. Building on Ms. Strassheim’s (8533) useful distinction between the scientists’ and regulators’ viewpoints I’d like to add some thoughts on the regulatory framework which may be apposite because as Mr. Linnestad said we discuss “impacts on biodiversity in the context of the three CBD objectives”.
1. As a preliminary remark we should be aware that the framework is, among others, the CBD, the Nagoya Protocol (NP), and the Cartagena Protocol (CP). The CP obliges to apply the precautionary principle. This means all of us should discuss on the basis of that principle (or propose to revise the CP), including also contributors from non-parties of the CP such as Australia, Canada, Russia, and the USA. The recurrently emphasised “evidence based regulation” should be reconsidered in that perspective. Moreover, the NP obliges to accept provider states’ regulation on access to their genetic resources (GR) and benefit sharing. I am not sure how non-parties of the NP (which include the same states as above) deal with such regulation. Anyway, contributors from non-parties should accept that the present discussion round is based on the NP.
2. In our round not much has been said about the 2nd CBD objective - sustainable use of biodiversity. This term opens up a broad span of possible definitions but “sustainable” at least limits uses in some way. Is the use for military purposes excluded? Does it warn against a vision of biodiversity as pure product of human design or “regenesis”, as some SynBio enthusiasts suggest (cf. Church & Regis 2012)?
3. A bit more but by far not enough has been said about the 3rd objective - the fair and equitable sharing of benefits with providers of genetic resources (GR). As some contributors pointed out a major challenge is here the fact that SynBio more and more works on GR related information rather than on GR as material. My suggestion: If a provider state by law or permit or contract stipulates a share in the benefits “arising” from the use of its GR this extends to benefits from products synthesized on the basis of information and without any material use of the GR. Provider states can also use their rights to determine access to their GR as a leverage to (by PIC and MAT conditions) control the use of information about their GR. This will be a serious challenge for bioinformatics (such as how to earmark information in data bases concerning origin and provider state conditions) and necessitates to discuss questions of exhaustion of benefit sharing claims (such as if the contribution of one gene gets lost in the noise of many other contributors). [By the way, my ceterum censeo in the ABS discourses is that the whole approach is overcomplex and should be replaced by a simple solution: free R&D plus a tax on monetary benefits arising from genetic resources, the revenue having to be given to provider states for biodiversity conservation. But as long as the CBD and NP are in force we need to act on their basis.]
4. The focus of the present discussion round has clearly been on the 1st CBD objective - the conservation of biological diversity. The rich and often controversial discussion could in some ways influence the regulatory approach. Here are some suggestions:
(a) The controversy about the definition and the risky kinds of SynBio can be read to reflect a need to readjust the scope of the regulation of biotechnology. The current trigger is whether the final product (for trade or release or containment) is a GMO/LMO. This covers most of SynBio, but not all, it does not cover certain new breeding technologies, and there may be some techniques that could be removed from regulation. A new approach could learn from from environmental impact assessment regulation (EIA) which combines lists of projects for obligatory EIA and criteria for screening further projects. A list of SynBio methods and products subject to obligatory in-depth scrutiny could be compiled plus criteria established allowing to screen further methods and products in terms of a need of in-depth scrutiny.
(b) Considering that the risk assessment methodology for GMOs is based on the comparison with familiar organisms new methods and tests must  be developed in reaction to the increasing artificiality of SynBio. Annex III of the CP needs to be adjusted accordingly.
(c) The criteria of risk management must be reconsidered. The focus on human health and the environment has been narrowed down to specific causal chains, such as from an LMO to non-target end-points.  It should be considered what strands of SynBio contribute to a kind of agriculture that “clears” huge areas and thus erodes biodiversity. It is no good objection to say that there are many other factors promoting industrialised agriculture. These factors must also be addressed, but in combination with adequate SynBio regulation.
(d) The question to what extent benefits of SynBio should be a regulatory yardstick needs further discussion. The European legal culture speaks against cost-benefit analysis (CBA), the US rather in favour. The CP acknowledges “socio-economic considerations”. My own position is against CBA, except for situations of residual risks that are often called negligible. Then “at least” a benefit should be shown, to be sure a use-value, not just monetary gains.

Firstly, I’d like to thank the opportunity to contribute, thank Casper for taking the challenge of moderating and thank the secretariat for setting this forum. I also took part in the AHTEG on SynBio that met in 2015. I am glad to see the level of interest in this forum.
Synthetic biology offers important benefits and potential solutions to a number of humanity’s most pressing ecological and animal/public health concerns. As raised by others (e.g. #8439, #8414, #8524, #8489, #8553,...) both potential positive and negative impacts are important and it’s wise to weigh them when taking decisions. It is worth to keep in mind, for instance, that some benefits might be greater than the potential negative impacts, that some adverse effects can be prevented with management and that technologies keep on being improved. Considering negative impacts and worst-case scenarios of Synthetic Biology is important to mitigate and prevent risks. This is part of appropriate and responsible Risk Assessment and Management. However, responsible management of risks is done in ways that we do not lose opportunities for improving human, animal and environmental health as well as gaining scientific knowledge. It should not compromise the development and legitimate application of synthetic biology and biotechnologies for our and the environment benefit. I agree with a number of postings that releases should be considered on a case-by-case risk assessment basis (e.g. #8430, #8427, #8447, #8538,...), these are done in a stepwise approach. I also agree that the existing risk assessment and management tools and practices (in line with annex III of the CPB) cover the current state of synthetic biology (e.g. #8563, #8533, #8550, #8663,…) and that monitoring of the developments should continue to make adaptations when and if needed.

My name is Simon Terry and I undertake research and analysis for the Sustainability Council of New Zealand.

In response to question 1 regarding adverse effects, a number of participants have counterposed: “yes, but is there anything new here?” (eg #8410, #8533, #8541, #8553).

That is another way of asking: “Does synthetic biology warrant change of the regulatory regime?”.  A key test of this for a regulator (and so the CBD) is whether the new tech can be used in ways that operate outside the assumptions and disciplines the current regime is founded on.

Gene drive is attracting considerable attention from the forum and it is an example of an application that straddles both the “adverse effects” and “what’s new” questions.  The “adverse affects” dimension of interest here is the potential ability of gene drive to eliminate a wild species in jurisdictions that it was not intentionally released into.  That is, an LMO designed to eliminate a wild species, and intentionally released in just one jurisdiction, but having its wider effect through a subsequent transboundary movement.

Such examples at the extreme are useful to stress test existing regulatory mechanisms (#8562, #8495).  While governance is not absent in this case, the current regime was clearly not founded with gene drive mechanisms in mind.

Such examples point to a review of whether the existing governance arrangements are fit for purpose (with respect to synthetic biology) and deliver on the objectives of the convention (see also #8581, #8495, #8451, #8437, #8529).

Dear All

I have seen a number of responses to my intervention that gave an example of gene disruption caused by introgression from genetically engineered rice. This was intended as a valuable example in discussing some of the risks associated with the release of SynBio organisms.

More detailed, my  intention was to give an example of gene disruption that was not only unexpected, but also had a significant impact in the organisms and, moreover, happened after many years of safety checks.

I gave the additional information that gene flow is known to occur between wild weedy rice and cultivated rice. This gene flow can, in fact, allow genetic conditions to persist and spread over longer periods of time.

Reading some of the contributions, I believe it is a valid argument to say that gene disruption observed in the specific case is associated with high fitness costs. Therefore, it might not be such a good example of new genetic information that can persist for a longer period of time.

However, I also believe that it is valid to point out that it is possible to introgress the genetic information into a variety that does not show the described effects of gene disruption  (research does indeed show that in many cases there are no observable effects). In this case, it could still be passed to weedy rice (without this being noticed as a potential hazard) and re-introgressed into other varieties cultivated in the region. These other varieties then might, or might not, show the effects detected in the Indian variety Swarna. Thus, the issue of fitness cost associated with the effects in the Indian variety Swarna does not necessarily limit the persistence of the relevant genetic information in the environment.

I believe the following conclusions can be drawn from this example:
> In many cases it is the genetic background that is decisive for the biological effects we see. And this issue would become much more pressing in regard to risk assessment if we were to start engineering native populations (with RIDLE, gene drives etc.).
> In general, we have to be aware of the limits of our knowledge and proceed with caution so that we can still withdraw the organisms from the environment, and prevent them from becoming persistent or from spreading uncontrollably.

In regard to regulation, we have to be aware that if there is a decrease in spatio-temporal control, there will be an increase in uncertainties in risk assessment: For example, risks with only a small probability of occurring within short period of time can become highly probable over a longer period of time.

Moreover, from the perspective of broader society, it would not be fair to burden future generations with the risks these present day organisms carry, since they could well be very quickly regarded as outdated or even risky from the point of view of future scientific evidence.

Therefore in conclusion, it appears that for us as a group, this is the right moment to discuss 'cut-off criteria' that allow us to identify those SynBio organisms which should not be released into the environment.

with kind regards 

Christoph Then

Dear All,
I want to first thank Mr. Linnestad for moderating such a robust discussion so far. My name is Todd Kuiken and I am a Senior Research Scholar with the Genetic Engineering & Society Center at North Carolina State University.  Prior to joining NC State I led the Synthetic Biology Project at the Woodrow Wilson International Center for Scholars. I have had the privilege to serve on the AHTEG on synthetic biology. Since I am joining the discussion towards the end and many issues have been raised and debated already, I would like to focus my intervention around technology trends, along with data and funding gaps as they relate to the guiding questions posed to the forum. 

Without minimizing the potential positive/negative impacts prior synthetic biology applications pose; the technology trend seems to be moving from “relatively” simple organisms (i.e. e-coli and yeast) to more complex organisms and environmental systems. Jim Thomas [#8437] provided a useful list and I would like to expand upon this using U.S. DARPA programs to illustrate this trend. Let me state clearly that I am not qualifying these DARPA programs as positive or negative. These programs provide examples of where the synthetic biology field may be moving towards and provide a window towards future applications that could have a direct impact on the objectives of the convention. While the phrase “synthetic biology” does not show up in DARPA budget documents until 2011, funding for “synthetic fuels,” “synthetic cells,” and “synthetic chromophores” begin to appear in 2008 and continued through 2010. Since then, DARPA has developed five programs, listed here in chronological order, that demonstrate the growing complexity of synthetic biology under development that could have a direct impact on the objectives of the convention.

The first program, called “Living Foundries”, launched in 2013. This program: “seeks to transform biology into an engineering practice by developing the tools, technologies, methodologies, and infrastructure to increase the speed of the biological design-build-test-learn cycle while significantly decreasing the cost and expanding the complexity of systems that can be engineered. The technologies and infrastructure developed as part of this program are expected to enable the rapid and scalable development of transformative products and systems that are currently inaccessible. Examples include novel materials, industrial chemicals, pharmaceuticals, and improved agricultural products (DARPA 2016a, Living Foundries Program).

The second program, called “Biological Robustness in Complex Settings” (BRICS), launched in August 2014. This program “seeks to develop the fundamental understanding and component technologies needed to engineer bio-systems that function reliably in changing environments. A long-term goal is to enable the safe transition of synthetic biological systems from well-defined laboratory environments into more complex settings where they can achieve greater biomedical, industrial, and strategic potential” (DARPA 2016b, BRICS).

The third program, “Safe Genes,” was announced in September 2016 and includes gene drives. This program is meant to “create biological capabilities that enable the safe pursuit of advanced genome editing applications.” According to the website: “Implementation of a ‘safety first’ approach to the development of next generation biotechnologies and genome editing tools and their derivative technologies (e.g., gene drives) will foster, and even accelerate, responsible innovation while mitigating the risk of unintended consequences. The Safe Genes program will provide new insights into what is possible, probable, and vulnerable with regard to genome editing biotechnologies and their derivative applications, create novel tools to enable predictable and reversible control of gene editors, and counter unwanted genome editing activity and outcomes” (DARPA 2016c, Safe Genes Proposers Day).

The fourth program, Insect Allies, was announced in November 2016. This program: “Aims to transform certain insect pests into ‘Insect Allies,’ by modifying insects to disseminate targeted genetic payloads to plant populations in order to protect crops from potential plant pathogens that are either naturally occurring or are intentionally designed and released to cause harm” (DARPA 2016d, Insect Allies). Finally, the fifth DARPA program, entitled “Ecological niche-preference engineering,” announced in 2017 centers around “the development of technologies that enable the genetic engineering of an organism's preference for a niche (e.g., temperature range, food source, and habitat). DARPA envisions creating genetic engineering strategies to control and alter the niche preferences of organisms to reduce economic, health, and resource burdens. A fundamental component of this work will be to expand our understanding of the genetic, epigenetic, and molecular contributors to the establishment of niche preference” (DARPA 2016e, Ecological Niche Preference).

Gene drives provide a good example of the gaps in knowledge that exist when attempting to address the guiding questions posed in this forum. The scientific data surrounding both the feasibility and ecological implications of gene drives is limited. Besides a few laboratory studies most of the scientific literature ascribing the benefits and risks of gene drives have been based on models and do not focus on ecological implications to address the broad scope of the questions posed in this forum [ (Hammond, et al. 2016) (Gantz, Jasinskiene, et al. 2015) (Gantz and Bier, The mutagenic chain reaction: A method for converting heterozygous to homozygous mutations 2015) (Di Carlo, et al. 2015) (Eckoff, et al. 2016) (Beaghton, Beaghton and Burth 2016) (Unckless, Clark and Messer 2017) (Reed 2017) (Esvelt, et al. 2014) (Noble, Olejarz, et al. 2017) (Noble, Min, et al. 2016) (Drury, et al. 2017) ].

As referenced by the U.S. State Department [#8550] the report produced by the Woodrow Wilson International Center for Scholars prioritized key research areas for government agencies, academia and industry to fund (Drinkwater, et al. 2014). Research areas include species for comparative research; phenotypic characterization; fitness, genome stability and lateral gene transfer; control of organismal traits; monitoring and surveillance; modeling and standardization of methods and data. The report says it is necessary to establish and sustain interdisciplinary research groups in order to conduct the research. Long-term support is also needed to address complex questions about how synthetic biology could affect the environment and overcome communication barriers across disciplines. However, there is still a significant funding gap that will need to be addressed in order to evaluate synthetic biology applications moving forward. An analysis of the U.S. federal research budget for synthetic biology suggests there is insignificant efforts to examine the ecological implications of these types of applications (Kuiken, U.S. Trends in Synthetic Biology Research Funding 2015).  Subsequently the U.S. National Academies report on gene drives concluded that there is insufficient evidence available at this time to support the release of gene-drive modified organisms into the environment (National Academies of Sciences 2016).

The Genetic Biocontrol of Invasive Rodents program is one example that may provide additional information for this effort. In full disclosure, I am a part of this program and therefor will not ascribe its potential benefits or risks, but simply as an example of a comprehensive program that is attempting to address the overarching questions posed to this forum in relation to synthetic biology and could have a direct impact on the objectives of the convention. The program is still in its early stages and there is limited data.  For an independent overview of the program, please visit: http://www.audubon.org/magazine/summer-2017/how-genetically-modified-mice-could-one-day-save
Finally, there were many interventions during this opening discussion around access and benefits sharing issues.  It is my understanding that there is an online forum and AHTEG being formed under the Nagoya Protocol to examine these issues, specifically in regards to digital sequence information. It may be helpful for the moderator to specify which topic areas will be addressed there.

References:

Beaghton, A., P.J. Beaghton, and A. Burth. 2016. "Gene drive through a landscape: Reaction-diffusion models of population suppression and elimination by a sex ration distorter." Theoretical Population Biology 108: 51-69.
DARPA. 2016. Defense Advanced Research Project Agency.
http://www.darpa.mil/about-us/about-darpa
DARPA. 2016a. “Living Foundries Program.”
http://www.darpa.mil/program/living-foundries
DARPA. 2016b. “Biological Robustness in Complex Settings (BRICS).” http://www.darpa.mil/program/biological-robustness-in-complex-settings.
DARPA. 2016c. “Safe Genes Proposers Day.” http://www.darpa.mil/news-events/safe-genes-proposers-day.
DARPA. 2016d. “Insect Allies Program.” https://www.fbo.gov/utils/view?id=40638c9e7d45ed8310f9d4f4671b4a7b.
DARPA. 2016e. “Ecological Niche Preference.” Young Faculty Award: DARPA-RA-16-63. https://www.fbo.gov/utils/view?id=c1540b48aa08624b27f4dc5e7cdf94fe.
Di Carlo, J.E., A. Chavez, S.L. Dietz, K.M. Esvelt, and G.M. Church. 2015. "Safeguarding CRISPR-Cas9 gene drives in yeast." Nature Biotechnology 33 (12): 1250-1257.
Drinkwater, K., T. Kuiken, Oye, K., S. Lightfoot, and J. McNamara. 2014. Creating a Research Agenda for the Ecological Implications of Synthetic Biology. Washington, DC: Woodrow Wilson Center. http://www.synbioproject.org/site/assets/files/1374/synbio_res_agenda1.pdf.
Drury, D.W., A.L. Dapper, D.J. Siniard, G.E. Zentner, and M.J. Wade. 2017. "CRISPR/Cas9 gene drives in genetically variable and nonrandomly mating wild populations." Science Advances 3 (5).
Eckoff, P.A., E.A. Wenger, H.C.J. Godfray, and A. Burt. 2016. "Impact of mosquito gene drive on malarial elimination in a computational model with explicit spatial and temporal dynamics." PNAS E255-E264. http://www.pnas.org/cgi/doi/10.1073/pnas.1611064114.
Esvelt, K.M., A.L. Smidler, F. Catteruccia, and G.M. Chruch. 2014. "Concerning RNA-guided gene drives for the alteration of wild population." eLife 3.
Gantz, V.M., and E. Bier. 2015. "The mutagenic chain reaction: A method for converting heterozygous to homozygous mutations." Science 348 (6233): 442-444.
Gantz, V.M., N. Jasinskiene, O. Tatarenkova, A. Fazekas, V.M. Macias, E. Bier, and A.A. James. 2015. "Highly efficient Cas9-mediated gene drive for population modification of the malaria vector mosquito Anopheles stephensi." PNAS e6736-e6743.
Hammond, A., R. Galizi, K. Kyrou, A. Simoni, C. Siniscalchi, D. Katsanos, M. Gribble, et al. 2016. "A CRISPR-Cas9 gene drive system targeting female reproduction in the malaria mosquito vector Anopheles gambiae." Nature Biotechnology 34 (1): 78-83.
Kuiken, T. 2015. U.S. Trends in Synthetic Biology Research Funding. Washington, DC: Woodrow Wilson Center. http://www.synbioproject.org/publications/u.s-trends-in-synthetic-biology-research-funding/.
National Academies of Sciences, Engineering, and Medicine. 2016. Gene Drives on the Horizon: Advancing Science, Navigating Uncertainty, and Aligning Research with Public Values. Washington, DC: The National Academies Press. doi:doi:https://doi.org/10.17226/23405.
Noble, C., J. Min, J. Olejarz, J. Buchthal, A. Chavez, A.L. Smidler, E.A. DeBenedictis, G.M. Church, M.A. Nowak, and K.M. Esvelt. 2016. "Daisy-chain gene drives for the alteration of local populations." bioRxiv. doi:http://biorxiv.org/content/early/2016/06/06/057307.
Noble, C., J. Olejarz, K.M. Esvelt, G.M. Church, and M.A. Nowak. 2017. "Evolutionary dynamics of CRISPR gene drives." Science Advances 3: 1-7.
Reed, F.A. 2017. "CRISPR/Cas9 gene drive: Growing pains for a new technology." Genetics 205: 1037-1039.
Unckless, R.L., A.G. Clark, and P.W. Messer. 2017. "Evolution of resistance against CRISPR/Cas9 gene drive." Genetics 205: 827-841.

Dear Colleagues,

This is Wei Wei from China. I worked on the biosafety issues of genetically modified organisms for nearly two decades and served as a member of the ATHEG group on risk assessment and risk management since 2008. I am very interested in studying the link between genetic engineering and synthetic biology on biosafety issues, and conducting related studies and aim to propose proper risk assessment and management strategies for synthetic biology.

In China, we are the first group to look into the biosafety issues of synthetic biology, and had published a report on the progress of this technology in China (http://www.sciencedirect.com/science/article/pii/S073497501100084X). Regarding of the risk assessment and potential effects of organisms derived through synthetic biology, the past experiences in the GM technology can contribute their help. I had provided this kind of description in a short opinion paper (https://www.omicsonline.org/open-access/biosafety-considerations-of-synthetic-biology-lessons-learned-from-transgenic-technology-2332-0737-2-1000115.php?aid=35307).

Sorry for the late participation in this forum. It takes a long time to read and study the huge information here provided by our distinguished colleagues. I appreciate all the valuable inputs and benefit a lot from this discussion. Thanks for this opportunity.

When we talk about the impacts of a technology, we may want to present both negative and positive ones. However, when we concern about biosafety issues and risks, it is not necessary to list all the positive ones. The technology shall have benefits to human people; otherwise, there is no reason to develop it.

As we are discussing under the CBD convention, thus all the impacts shall be related to the main contents of biodiversity conservation, including sustainable development and two other protocols (biosafety and ABS) etc. I think all the pointed proposed by our colleagues are valid, especially those by [#8413] and [#8437] and many others. I agree to our colleagues that the most risky effects line in the new emerging genome editing tools (including gene drive), off-targeted effects and DNA synthesis using extensive sequence information available.

With best wishes

Wei

Dear All – I am Kelebohile Lekoape with experience in Biosafety and Regulatory Affairs. Thank you for the opportunity to contribute to this on-line forum.  I will answer the specific questions raised by Casper.
1. What are the potential negative impacts, including unexpected and significant adverse effects, of the most recent technological developments in synthetic biology on biodiversity and the three objectives of the Convention?

I am of the opinion this question has already been addressed, but am wondering if the emphasis on potential negative impacts is to differentiate this from previous discussions; if so, I would suggest examples relating to specific technologies should be provided in order to engage this topic in a meaningful way.

2. What research and cooperation activities are being conducted on the possible benefits and potential adverse effects of organisms, components and products of synthetic biology on biodiversity to fill knowledge gaps and identify how those effects relate to the objectives of the Convention and its Protocols?

In South Africa, a number of research institutions (public and private) collaborate among themselves and with international organisations on research activities that mainly involve the use of gene editing and genetic modification technologies.  These operations are conducted in laboratory settings where the appropriate risk management practices are applied.  The risk assessments and case-by case evidence base have not demonstrated the need for an amendment in the legislative arrangements currently in place.  To my knowledge none of these research activities specifically investigate the impact such technologies may potentially have on biodiversity.

Dear forum participants,

In topic 1 posted by our moderator we are asked to review recent technological developments within the field of synthetic biology to assess if the developments could lead to impacts on biodiversity and the three objectives of the Convention, including unexpected and significant impacts.

On the question of recent technological developments many on the forum have identified a range of biotechnological tools and developments, predominantly research applications, that are qualified (without complete consensus) as belonging to the field of synthetic biology.  I believe that what we have been describing so far on this forum is the continuous progress in the field of biotechnology and the convergence of scientific disciplines to address new scientific questions as well as to re-examine existing data. It is not possible to classify scientific progress as risky per se, or as having a specific  impact (positive, negative, or neutral) unless we consider on a case-by-case basis  the applications developed with this knowledge and in comparison with existing alternatives.  Furthermore such comparison should be based on evidence and supported by high quality scientific data.
The example below  is a good illustration of the difficulty that often occurs when interpreting partial, incomplete or badly designed experiments and why it is so important to have sound scientific data in support of risk assessment.
The article of Mesnage et al.,  2016, Scientific Reports 6, 37855 was quoted in this forum as an example providing evidence on “unintended effects” in an LMO. The European Food Safety Authority [EFSA Volume 14, Issue 6, June 2017, http://onlinelibrary.wiley.com/doi/10.2903/sp.efsa.2017.EN-1249/abstract ] has reviewed the above article and concluded “…that there are severe shortcomings in the experimental design, as well as uncertainties on the suitability of the test material as described by Mesnage et al. (2016).  Furthermore, the interpretation  of the results is incomplete, since the authors did not take into account natural variability of the endpoints analysed. Uncertainty remains with regard to potential contamination of the study materials by pathogens and the related confounding  effects on the study results. The publication  by Mesnage et al. (2016) does not reveal any new   information  that  would  invalidate  the  previous  conclusions  on  maize  NK603  made  by  the EFSA GMO Panel. Therefore, EFSA considers that the previous risk assessment conclusions on maize NK603 remain valid and applicable.”

Best regards,
Ana

Dear All

I have seen several comments regarding risk assessment that make a comparison between the genetic changes introduced by genome editing and those due to spontaneous mutations.

Some experts have already responded by saying that there are checks and balances in normal gene regulation and epigenetics which may, to some extent, be able to control spontaneous changes in the genome. But gene editing can circumvent these mechanisms. So to me, there seems to be a relevant difference between effects caused by spontaneous mutations and those triggered by genome editing.

There is an additional point we should consider: even if no additional gene material is inserted, as  in the case of the non-browning mushrooms produced by  Penn State University, the (on target) pattern of genetic changes introduced by CRISPR-Cas (and other methods of genome editing) is vastly different in comparison to spontaneous mutations. Using CRISPR Cas technology resulted in parallel changes in the genome of the mushrooms to block the gene function that naturally causes browning. Such a specific pattern of parallel changes in the genome would not occur spontaneously. However, with CRISPR-Cas technology several parallel changes in the genes are typical and often unavoidable: These nucleases cut at all the sites in the genome where the respective gene sequences are located. For example, specific genetic information is often located at several sites in the genome of crop plants.

So a comparison between random, spontaneous mutations (or random mutagensis) and the changes introduced by genome editing (on target or off target) seems to have only very limited benefits when it comes to risk assessment.

with kind regards,

Christoph Then

Dear Casper
Dear Participants

Greetings from Iran!
My name is Behzad Ghareyazie (PhD Genetics). I am Cartagena Protocol on Biosafety National Focal Point of the Islamic Republic of Iran. I have been involved in CBD, CPB and Nagoya Protocol negotiations during the last 20 years.
I have been carefully reading and reviewing the comments and submission by the other respected participants of the forum during the last two weeks together with a team of experts from different stakeholders working hand-in-hand with the Iran CPB NFP.
As soon as we came back from Cancun (COP 13/MOP 8), we established a committee on SynBio to get prepared to contribute to this forum and the following AHTEG meetings.
With due respect to those who are dissatisfied with the operational definition of synbio as the outcome of the AHTEG, agreeing with the fact that the definition does not “define” anything more than what one can understand from the term “Synthetic Biology”, I wish to comply with the command made by Casper (#8377), asking to refrain arguing about the definition.
I do however wish to clarify that our understanding of the definition is that synthetic biology is only “a further development and new dimension of modern biotechnology”. We failed to get any example of synthetic biology that could fall outside the definition of modern biotechnology (as defined in CPB). Therefore, I wish to support Ms. Motlalepula (#8516) and those who supported her contribution. Therefore, I would like to emphasize that we already have a very well negotiated “Cartagena Protocol on Biosafety” for responsible deployment of Modern Biotechnology, including experienced methodology for risk assessment.
We are deeply concerned about opening new line of negotiation for synthetic biology, which is no different from modern biotechnology as mentioned above. Even the participants of the forum who are requesting for more stringent restriction on the technology (e.g. Mr. Taye #8385 and #8402), have the same understanding that synbio is part of modern biotechnology. That is why the concerns that they have raised in the forum are the same as the concerns raised during the last 25 years about the potential negative impact of LMOs on the conservation of biological diversity. The raised concerns include (but are not restricted to) the distribution of resistance genes (e.g. #8409 and #8418), invasiveness (e.g. #8404), unexpected outcomes and overcoming the natural barriers (e.g. #8392, #8395 and #8436), gene scape and horizontal gene transfer (e.g. #8399 and #8408), effect on non-target organisms (e.g. #8399, #8400 and #8403) and uncertainty (e.g. #8501 and #8523).
On the other hand the emphasis on the benefits of synbio and the necessity for assessing the potential risks on the “case-by-case” basis (e.g. #8409, #8410, #8418, #8408, #8417, #8435, #8447 and #8528) is in line with CPB again.
Synbio can be used to synthetically produce products that are currently produced or extracted from plants and/or animals. A good example is the production of synthetic Saffron which is the main source of livelihood of tens of thousands of Iranian Saffron producers who produce more than 90% of the world Saffron. This may not only have any negative effect on the conservation of biological diversity, but may even have positive effects since the saved water can be used for the environment. However, it will surely have negative socio-economic impacts, local demographic displacement of the farmers and even will raise ethical issues. Even saying so, this issue can be accommodated very well under the socio-economic considerations of the Cartagena Protocol on Biosafety (Article 26) and the following discussions in the area.
Supporting the view and information posted by Andrew Roberts (#8410) seconded by #8427, I would like to emphasize that we should not consider the risk of any modified organism including synbio products Per-se. we should rather emphasize on the new risk(s) posed by synbio that is not posed by other organisms sharing the same phenotype. For example why should the HGT be considered as a concern from an LMO or synbio product resistant to an insect or tolerant to a certain herbicide but not considered a concern when exactly the same phenotype is transferred though HGT from a non-LMO/non-Synbio organism?
Supporting the view expressed by Luck Alphey (#8415) on HGT and effect on NTOs. I do see similarities between the discussions on CPB during the last decades and synbio discussion today. On one hand there are participants who are ideologically against the technology and the concerns they raise are usually hypothetical and speculative and perhaps never “evidence-based”. They even call for moratorium. On the other hand there are participants who emphasize on the application of synbio beneficial for biological diversity (e.g. #8549). Therefore, I wish to conclude that:
1) synbio does not pose any new threat in comparison with what is posed by modern biotechnology.
2) Responsible deployment of the technology and gaining benefit from it should be facilitated and access of the developing countries to its benefit should be guaranteed.
3) CPB is perfectly suitable for regulation of synbio and the risk assessment principle contained in CPB is enough to accommodate the raised concerns.
4) I wish to recommend that any further discussion on synbio is mandated by CBD to CPB during the next COP.

The valuable contribution of Gerd Winter [#8581] resembles the position advanced by the Peruvian Society of Environmental Law (SPDA) in various fora and refereed publications. Participants should take note that the two are not isomorphic. The distinctions are worth highlighting as they contrast economic thinking with the lack thereof.  Item (3) of Prof. Winter’s posting invites careful analysis.

Prof. Winter refers to “digital information” and not “natural information”. However, “digital” is merely the medium of transmission of natural information; isolation of “digital” from other media of transmission would exclude those other media [#8573]. Under Prof. Winter’s suggestion to include “digital information” with “genetic materials” for the purposes of ABS, lawful avoidance could proceed through those other media. Thinking economically, incentives would emerge to access natural information through media that are not material or digital (SPDA Submission of Views for Notification 2017-37, titled ““Unpacking ‘Digital Sequence Information on Genetic Resources’: Scaffolding of Errors to Preserve a Category Mistake’”, forthcoming 2017).

Prof. Winter’s further argues that “Provider states can also use their rights to determine access to their GR as a leverage to (by PIC and MAT conditions)”. Inasmuch as  natural information is the object of access for R&D, shareholder-held firms will have a fiduciary responsibility to source the medium (genetic material) where it is cheapest. So one would not expect much “leverage (by PIC and MAT conditions)”. Most natural information is transboundary and “jurisdiction shopping” will guarantee that royalty payments are so low that firms will not dare publish them for fear of “getting RAFI’d” (McManis, 2004, p. 460). The SPDA explains the economics in its submission of a new and emerging issue for COPIV (https://www.cbd.int/emerging/).

Prof. Winter’s statement “[t]his will be a serious challenge for bioinformatics” would also not be true under the position advanced by the SPDA.  Many of the building blocks of synthetic biology (see LEGO analogy in [#8434]) would have entered the public domain with the expiry of patents over the value added. For pure synthetic biology, no ABS requirement would be triggered.  

A critically important distinction between the position published by the SPDA and that of Prof. Winter lies in the following statement: “The whole approach is over[ly] complex and should be replaced by a simple solution: free R&D plus a tax on monetary benefits arising from genetic resources, the revenue having to be given to provider states for biodiversity conservation.”  Albert Einstein famously said “Everything should be made as simple as possible, but not simpler.”  Prof. Winter’s suggestion of a “tax on monetary benefits arising from genetic resources” is “too simple”. An alternative which is simpler than the “over[ly] complex” MTAs but not “too simple”  goes under the nomenclature “bounded openness”.

Political scientist Chris May (2010) coined the term and described what he meant in the sphere of informatics and copyrights. However, Prof. May did not define “bounded openness”, which is a strategy we can all appreciate! Inspired by May’s description, the SPDA defined the term thus:

Bounded openness: Legal enclosures which default to, yet depart, from res nullius [property of no one] to the extent the departures enhance efficiency and equity, which must be balanced when in conflict (Peruvian Society of Environmental Law, 2016, p2, fn2)


Under “bounded openness” as the modality for the Global Multilateral Benefit-Sharing Mechanism of Article 10 of the Nagoya Protocol, natural information would flow unencumbered for R&D. There would be no requirement for prior informed consent. Disclosure of utilization would occur with the application for intellectual property protection. Only for commercial successes  would the levy of a royalty be triggered. Because of highly diverse utilizations, the royalty would not be the same flat percentage across industries and types of intellectual property.  The mechanism of how the royalties would be designed was introduced in “Genetic Resources as Natural Information” (Ruiz Muller, 2015) and elaborated in the “Bounded Openness as the Modality for the Global Multilateral Benefit-Sharing Mechanism of the Nagoya Protocol” (Vogel et al, forthcoming 2018).

The contrasts continue. Prof. Winter’s statement that  “the revenue having to be given to provider states for biodiversity conservation” ignores fungibility, which is a common mistake by non-economists. Many activities for biodiversity conservation are already publicly funded; by switching the financial source to ABS, conservation would not necessarily be more funded. Moreover, in many megadiverse countries, the opportunity costs of land use is the driver for extinction (the H in HIPPO, an acronym which needs no elaboration). Paradoxically, what often needs to be done is to not do something: not to open new roads into the Amazon; not to dam rivers; not to drain wetlands; and so on. Bounded openness over natural information would remit royalty payments to countries based upon estimates of the habitat of the species that contain the utilized natural information that was commercially succesful. So, if habitat dwindles due to changes in land use, so would the proportional ABS claim of the country that shares the natural information.

Finally, Prof. Winter’s closing sentence in item (3) is as dispiriting as it is seemingly true.  “But as long as the CBD and NP are in force we need to act on their basis.” We must remember the well documented history of the drafting of the CBD, which “suffer[ed] from basic conceptual and drafting deficiencies. The structure of the negotiations, the haphazard way in which crucial issues were considered, and the pressures of time...which should cause distress for international lawyers and policy-makers” (Chandler 1993, p. 174). To counterbalance the deficiencies, the CBD was set up as a framework convention. Thank goodness.

With the political will of the COP, the flaws of the CBD and its subsequent Protocols can be corrected. The foundational error in defining a “genetic resource” as “material” in Article 2 must be discussed if the three objectives are ever to be achieved.

Chandler, M. (1993) “Biodiversity Convention: Selected Issues of Interest to the International Lawyer”. Colorado Journal of International Environmental Law and Policy 4(1): 141-175.

May, C. (2010) The Global Political Economy of Intellectual Property Rights, 2 edn, Routledge, London.

McManis, Charles R. (2004). “Fitting Traditional Knowledge Protection and Biopiracy Claims into the Existing Intellectual Property and Unfair Competition Framework”. Chapter 12 in Intelletual Property and Biological Resoures, Burton Ong, ed (Marshall Cavendish Academic).

Ruiz Muller, Manuel. (2015) Genetic Resources as Natural Information: Policy Implications for the Convention on Biological Diversity (London: Routledge).

Peruvian Society for Environmental Law (SPDA) (2016) ‘Submitted view for the Updated report and synthesis of views in response to paragraph 7(b) of Decision XII/24; and Report of the Meeting of the Ad Hoc Technical Expert Group on Synthetic Biology’, http://bch.cbd.int/synbio/peer-review

Vogel Joseph Henry,  Klaus Angerer, Manuel Ruiz Muller and Omar Oduardo-Sierra, (forthcoming 2018) “Bounded Openness as the Global Multilateral Benefit-Sharing Mechanism for the Nagoya Protocol”. Chapter 26  in Charles R. McManis and Burton Ong (eds) Routledge Handbook on Biodiversity and the Law (London: Routlege).

Dear forum colleagues, dear Casper,

First my sincere thanks to Casper for agreeing to moderate this on line discussion, and for his characteristic kindness in thanking people for the “wide array of interesting views”, despite that he had actually - and rightfully - urged everybody to stick to the three questions he posed at the beginning and to be concise and focused.

As we have all seen, many of the over 200 posts do not focus on the three questions. Moreover, quite a number of posts discussed aspects that are not specific to Synthetic Biology (e.g. off target mutations, I fully agree with the sublime post of Tizard on this), and quite a number of posts discussed technical developments or applications that are not Synthetic Biology in themselves (e.g. CRISPR). As one colleague said, it is as if every new technology that has been developed after the adoption of the Cartagena protocol is lumped in this discussion, regardless whether it has anything to do with Synthetic Biology. What confounds matters further is that in several posts that neither addressed Synthetic Biology as such nor aspects specific to Synthetic Biology, some challenging concepts were presented that in turn provoked reactions from experts in the respective fields.

I very much hope that Casper and the Secretariat will be able to keep the summary of this debate limited to the posts that actually did address the three questions and that did focus on Synthetic Biology as such or aspects specific to Synthetic Biology.

With that introduction, let me address the 3 questions Casper listed: 

1) What are the potential negative and positive impacts of the most recent technological developments in synthetic biology on biodiversity and the three objectives of the Convention? (I very much agree that this question should also address positive impacts). 2) What research and cooperation activities are being conducted on the possible benefits and potential adverse effects of organisms, components and products of synthetic biology on biodiversity to fill knowledge gaps and identify how those effects relate to the objectives of the Convention and its Protocols?

As regards potential negative impacts: whether certain applications of synthetic biology would pose risks to the conservation and sustainable use of biodiversity cannot be answered in a general fashion. Yet, we should recognise that the current and foreseeable applications of synthetic biology will fall under case by case risk assessments of the existing biosafety systems such as the Cartagena Protocol. The risk assessment under the Cartagena Protocol instructs to include relevant data of the recipient or parental organisms, the insert and the donor, the transformation system, the intended use and the potential likely receiving environment.

With regard to potential positive effects of applications of synthetic biology to the conservation and sustainable use of biodiversity: as was illustrated at some of the side events at COPMOP2016, the range of applications and potential positive effects is so vast and diverse that it could fill a whole series of online discussions.

For example, the COPMOP2016 side event held by iGem students active in synthetic biology discussed topics such as producing biofuels from air, using plant pigments to create more efficient and sustainable solar cells, desalinizing water with bacteria, improving the production of graphene, a promising material in the biomedical and energy areas, and using carbon nanotubes to make scientific research easier to carry out in poor communities. These cases represented just a tiny, tiny fraction of the multiple lines of research in many research institutes, and it would be very beneficial if some of the online discussions could be dedicated to discussing some of these examples in detail.

Looking forward to the next round of debates!

Piet

"Dear Casper, dear all,

Thank you very much for moderating Casper and thank you all very much for the interesting and informative discussion. I have only this evening had the possibility to read all the comments posted to the forum.
My name is Marja Ruohonen-Lehto and I have worked over 20 years on LMO risk assessment and biosafety issues. I have participated in the Cartagena negotiations and followed SynBio both in SBSTTA and in COP negotiations. I do not have a lot to add - many excellent contributions have already covered a lot of issues. I am happy that we moved forward from the definition issue - we should use the AHTEG developed definition during our present discussions and when getting ready for the AHTEG and SBSTTA in December 2017. Our focus should at this point be on possible negative effects of SynBio on biodiversity and  the three objectives of the Convention. This discussion should, when possible use specific examples of SynBio applications. Finland has sent a short national reply on research conducted in Finland. Finally, several good suggestions were posted on "other recent technological developments that have taken place within the field of synthetic biology that need to be considered in this discussion?". Here I would like to just mention gene drives and maybe especially in insects - due to present research on mosquitoes.
Thanks a lot once more for good discussions - I look forward to our next rounds of discussions.
All the best,
Marja Ruohonen-Lehto

Dear forum participants,

I would like to address points raised in recent posts related to uncertainty and comparators in risk assessment. These points have already been discussed previously in the synthetic biology online discussions in 2015, as well as in the Open-ended online forum on risk assessment under the Cartagena Protocol who considered synthetic biology risk assessment in 2016 (see: http://bch.cbd.int/onlineconferences/2014_2016period.shtml). These have arisen due to the concerns of some that increasing complexity of genetic modification correlates with increasing uncertainty and decreasing availability of appropriate comparators. Generally speaking, it is feasible that as the characteristics of an LMO become more familiar to regulators, the risk assessment should become less complex, and that new and different types of LMOs may present less familiar characteristics – however, today’s “familiar” LMOs were once also “new” and “complex”. For both of these issues, regulators participating in the risk assessment online forum did not consider that new and more complex LMOs presented new risk assessment challenges that could not be managed using existing approaches.

In regard to uncertainty, it is stated in a recent post that the absence of evidence for a negative effect is not in itself evidence for the absence of a problem. The principles of Annex III include a broader statement on uncertainty, where “lack of scientific knowledge or scientific consensus should not necessarily be interpreted as indicating a particular level of risk, an absence of risk, or an acceptable risk”. In practice, to manage uncertainty, regulators may take a range of actions, e.g. request additional investigations in order to improve scientific knowledge relevant to the evaluation of risk, decide to release the LMO with risk management measures or monitoring requirements, or decide that the risk assessment cannot be completed and refuse release of the LMO until such time that there is improved scientific knowledge. The same approach will continue to apply to LMOs with increasingly complex genetic modifications. Claims that technology and products are developing so fast that there will not be sufficient time for regulators to properly conduct risk assessments do not reflect reality. A lack of scientific knowledge or understanding on the part of regulators will simply result in an increase in the time taken for risk assessment – it is not plausible that regulators will proceed with approving products for release into the environment if they cannot complete the risk assessment. In the case of gene drives for malaria control, the uncertainty is around the specific application rather than the complexity of the genetic modification. However, the same general approach will apply, and in this discussion the gene drive scientific community has made it clear that they are actively working to improve scientific knowledge.

Annex III also includes assessment of the LMO “in the context of the risks posed by the non-modified recipients or parental organisms”. In practice, risk assessment of LMOs has used such a comparative approach, e.g. LM crops are typically compared to the original non-modified line/variety as this is the closest genotype to the LMO. However, such a narrow approach is not mandated by Annex III, and alternatives (e.g. additional lines, other LMOs) may be more appropriate on a case-by-case basis. Concerns about appropriate comparators are due, in part, to speculation on the types of LMOs that may be developed in the future which tend to overstate the current state of the science and technical capabilities. The closest we have come to “creating new life” is chemically synthesizing genomes for microorganisms, where the host cell is the existing living organism, and arguably this will remain the case for the foreseeable future. At the very least, where the host is an existing species, that species (and/or higher taxonomic levels) provides a comparator that can be used in risk assessment. Such an approach has already been reported by regulators for some modified microorganisms (e.g. Australia and Brazil – risk assessment online forum).

It is important to remember that risk assessment methodology is not fixed in time or adapted only to “familiar” LMOs. A fundamental principle of Annex III is the case-by-case approach – this allows for flexibility depending on the organism, its intended use and receiving environment, and recognizes that the required information may vary in nature from case to case.

Dear all,
my Name is Birgit Winkel and I work for the German federal agency for nature conservation as a risk Assessor for more than 14 years.
I read the post with an interesst and just picked some to reply on their Statements:

It may be right that a lot of appplications of organisms produced by SynBio are not knew, but we habe to take into account that the Quality of the influence is deaper.

To post 8449: It is right that prediction of future applications of a new Technology is almost impossible, but it is necessary to be Aware of possible Impacts to be ready to Access their risk! That is the reason why SynBio is such a new and emerging technic on which we Need to Focus our assessment.
To post  8460: No matter if SynBio may have potential benefits that should never hinder to have a Close look to the potential risks. This Technology is fast developing and the potential changes can be of a new Quality. We Need to know which Tools we have, to be able to perform an appropriate risk assement.
To Posts 8493 and 8533: In my opinion even if all existing Syn Bio organisms can be called LMO/GMO existing risk assessment Tools are not enough to assess organisms that are not comparable with living organsims, because they differ to much.
Our current risk assessment is build on the comparative Approach that clearly does not work any more, if the change is so deep that the perceived organism is ot comparable to any existing organism.

Regards
Birgit