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Topic 5: Consider whether any living organism developed thus far through new developments in synthetic biology fall outside the definition of living modified organisms as per the Cartagena Protocol

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Welcome message - synbio forum - topic 5 [#9602]
Dear Participants,

Welcome again, this time to discuss topic 5. The aim of this discussion is to provide examples of synthetic biology organisms (if any) that may fall outside the definition of living modified organisms as per the Cartagena Protocol. As you know, the COP has requested the AHTEG on synthetic biology to take into account also the work under the Cartagena Protocol and have called for a coordinated approach between the CBD and its Protocols.

I would like to remind you that the Ad Hoc Technical Expert Group on synthetic biology in 2017 concluded that "most living organisms already developed or currently under research and development through techniques of synthetic biology, including organisms containing engineered gene drives, fell under the definition of LMOs as per the Cartagena Protocol." ( https://www.cbd.int/doc/c/aa10/9160/6c3fcedf265dbee686715016/synbio-ahteg-2017-01-03-en.pdf , please see page 5 of the report) Has this picture changed?

Please bear in mind that Parties may have different definitions on LMOs according to their national legislation, and also, that genome edited organisms may be regulated differently.

The Cartagena Protocol on Biosafety has defined living modified organisms as

"...any living organism that possesses a novel combination of genetic material obtained through the use of modern biotechnology”.

Furthermore, modern Biotechnology is defined by the Cartagena Protocol as

"…the application of:

a. In vitro nucleic acid techniques, including recombinant deoxyribonucleic acid (DNA) and direct injection of nucleic acid into cells or organelles, or

b. Fusion of cells beyond the taxonomic family, that overcome natural physiological reproductive or recombination barriers and that are not techniques used in traditional breeding and selection”.

I finally take the opportunity to kindly suggest that you reply to this message when making interventions. This will facilitate the subsequent work on summarizing the information and writing the report. Please also refer to the # of a particular post if you are responding to it.

I am hoping for a fruitful discussion!

Regards,

Casper Linnestad
posted on 2019-03-18 01:02 UTC by Mr Austein McLoughlin, Secretariat of the Convention on Biological Diversity
RE: Welcome message - synbio forum - topic 5 [#9605]
Dear Colleagues,

The issue of any living organism developed thus far through new developments in synthetic biology fall outside the definition of living modified organisms as per the Cartagena Protocol had been discussed extensively under Topic 3 of the previous on-line forum (https://bch.cbd.int/synbio/open-ended/discussion_2017-2018/#topic3). I think two cases need to be looked at as possibly falling outside of the definition of LMOs:

1. Organisms whose genomes had been edited without the use of nucleic acids using only protein reagents (Mega nucleases, ZFN, TALEN) introduced into the cell.
This issue has also been raised in the Austrian submission of information (http://bch.cbd.int/database/attachment/?id=18947) and I have some vague recollection that it was mentioned in the previous on-line forum under Topic 3. In this case the proteins act as sophisticated mutagens and the organisms probably will not be LMOs as no in vitro nucleic acid techniques have been used. On the other hand I doubt that such organisms will fall within the scope of Synthetic biology.

2. Virus-like macromolecular assemblies.
This issue has been raised in the Bulgarian submission of information (http://bch.cbd.int/database/attachment/?id=18988). It was demonstrated that it is possible to create virus-like entities consisting of computationally designed icosahedral protein assemblies capable of packaging their own full-length mRNA genomes (Evolution of a designed protein assembly encapsulating its own RNA genome, Butterfield et al., Nature 552 (7685), 415-420). The created macromolecular assemblies are certainly obtained through the use of modern biotechnology. Those assemblies do not have all functions required for a complete viral life cycle and are not capable of transferring or replicating their genetic material. Thus they cannot be considered living organism, but it can be imagined that similar non-biological synthetic assemblies might be developed in the near future that will be capable to enter and replicate inside cells and be even more similar to naturally occurring viruses.

Best Regards,
Nikolay
posted on 2019-03-18 08:37 UTC by Mr. Nikolay Tzvetkov, Bulgaria
RE: Welcome message - synbio forum - topic 5 [#9606]
Dear Casper & colleagues

As indicate in an earlier intervention I am seeking to better define Synthetic Biology and would like to make the following suggestion:

The current broad ‘definition’ (really a description, as per Wei Wei #9465) of Synthetic Biology (Synbio) overlaps and includes genetic modification. This continues to cause confusion - we can reference a great number of messages in this forum already that refer to this confusion.

The confusion is caused - in my opinion - because we have not clarified the ultimate purpose of this initiative.  In my opinion there are/could be two main purposes:
1. We want to be aware of new techniques and tools that are relevant to manipulating the genome (this purpose therefore includes LMOs within Synbio), and
2. We are aiming (ultimately) to provide regulatory guidelines -as already provided for LMOs- for Synbio.

For the former, we need to clearly acknowledge that the new techniques and tools may be used to different effect, and it is the effect/product, not so much the techniques/tools, that would need to be examined for regulatory implications. The pragmatic definition of Synbio under purpose 1 is all techniques/tools that can be used to manipulate the genome (without consideration of regulatory implications).

To me as a regulator it would be more satisfying to focus on purpose 2: we already have regulatory guidelines for LMOs, and we need to focus our attention on ‘beyond’ LMOs.  To this end we need to differentiate Synbio from LMOs.

Thus a pragmatic definition of Synbio under purpose 2 would be genetic engineering that extends beyond mere modification of the genome of organisms.

How practically can we differentiate?  Under current regulation, LMOs are compared with their ‘conventional’ (ie. not genetically modified) isolines or near isolines, ie. the principle of Substantial Equivalence generally applies.

What is needed is consideration of genetic engineering BEYOND what is ‘mere’ modification, ie beyond what is currently regulated under the principle of substantial equivalence.

The issue of regulation of Synbio thus becomes apparent only when there is no near isoline because there are at least substantial genetic differences from the conventional counterpart.  We would then need to define what is meant by substantial.

This could include (a) genetic-based SYSTEMS (complex metabolic functions) originating from one or more unrelated species, or designed de-novo; (b) gene-based proteins or functions that do not occur in nature; (c) the organism comprising engineered genetic-based metabolic pathway(s) - not just protein expression - that results in a different purpose for the pathway; or (d) because the impact of the organism has a significant divergence (in terms of impact) from the non genetically engineered organism.

The first three types (a-c) are aimed at identifying the ‘substantial-ness’ of the genetic change, and the last (d) is to cater for substantial change in impact.

I would think that in our postings, we should then state whether we are addressing purpose (1) or purpose (2).
posted on 2019-03-18 09:06 UTC by Mr. Ben David Durham, South Africa
RE: Welcome message - synbio forum - topic 5 [#9608]
I want to thank Maria, Casper and all participants for the interesting discussion on topics and the prolific literature to enrich the discussion. As has been stated in several posts from previous topics, there is a confusion on a clear definition of what synthetic biology is and what its boundaries are.
Also, the Cartagena protocol on biosafety defines LMOs as "...any living organism that possesses a novel combination of genetic material obtained through the use of modern biotechnology” and Modern biotechnology is also considered the application of “In vitro nucleic acid techniques, including recombinant deoxyribonucleic acid (DNA) and direct injection of nucleic acid into cells or organelles”.
What is considered a novel combination?
One may interpret as if any living organism developed through i.e. genome editing even the ones that are proved to be free of any transgene or of any unintended undesirable effects (off-targets) or that were indistinguishable from traditional breeding were failing into this definition of LMOs.
There are several posts on topics 3 and 4 that are relevant to this discussion on topic 5, and I will just quote some of them:  
[#9588] Piet van der Meer (topic 4): “while SynBio can include the use of genome editing, not every application of genome editing results in SynBio. Likewise, not every application of genome editing results by definition in an LMO”.
Felicity Keiper #9572 (topic 3): “Genome editing represents a broad category of enabling tools that can be applied to achieve various outcomes in plants, with some outcomes comparable to existing biotech (LMO/GMO) plants, and others comparable to plants developed with conventional breeding tools. Therefore, current and foreseeable applications of genome editing in plant breeding do not result in crops that are fundamentally different from those currently commercialized”
Louisa Matthew #9553 (topic 4): “While gene editing is one of the many tools used in synthetic biology, I do not consider that every application of gene editing results in a synthetic biology organism. As has been stated elsewhere, it is the outcome, not the tools used, that determines whether or not a particular application is synthetic biology.”

While i.e. CRISPR/associated endonuclease is used for different Synbio applications, using it to generate an improved organism does not necessarily make it a Synbio product, which favors a product-based oversight and a case-by-case responsible and scientifically-based dependent approach. As stated by other members under previous topics, regulation should protect health and environment but avoid unjustifiable barriers to innovation. Given all new and coming possibilities, probably a more accurate definition on LMO under the Cartagena protocol as well as a clear Synbio definition may help overcome hurdles on innovation while maintaining a responsible scientifically based oversight.

I would like to cite some examples that may be considered non LMOs under new developments and / or that provide new alternatives to reduce unintended effects of genome editing (i.e. off-targets), useful for the same purpose. 

One of them is reviewed by Chilcoat et al. (2017) that pointed out examples of new plant varieties indistinguishable from those that could be obtained by traditional breeding which can be, but not limited to, the generation of null alleles using template free genome editing (null alleles are spontaneously generated in nature, sweet corn is an example) or the precise transfer of a single allele from one variety to another of the same species or a close crossable wild relative harboring i.e. a disease-resistant allele, which by traditional breeding may require an expensive, imprecise and time consuming backcrossing process (https://www.ncbi.nlm.nih.gov/pubmed/28712499). These examples may be considered non LMOs, which will encourage the development of precise and useful changes of locally adapted varieties, more affordable to i.e. non-profit organizations that work for the benefit of local communities.

Liang et al (2017) stablished an efficient and specific CRISPR/Cas9 RNP mediated genome editing method for bread wheat suggesting that the use of CRISPR/Cas9 RNPs will greatly aid specific and clean genome editing in a wide range of plant species as plants do not have integrated transgenes  (https://www.nature.com/articles/ncomms14261). This kind of applications of genome editing would probably be not be considered as an LMO if is free of transgenes and unintended effects.

Metge-Sprink et al (2019) reviewed technical and regulatory perspectives pointing out that off-targets play a minor role in DNA-free approaches (https://www.frontiersin.org/articles/10.3389/fpls.2018.01957/full).

Another paper by Tang et al (2018) compares mutations produced by CRISPR/endonucleases with conventional technologies exemplifying a way to overcome concerns of genome editing in plants. It showed that off-target mutations using Cas9 and Cpf1, two leading CRISPR genome editing systems, in rice are largely negligible when compared to spontaneous mutations that arise from seed propagation or mutations caused by tissue culture and Agrobacterium infection in edited plants (https://www.ncbi.nlm.nih.gov/pubmed/29973285).
posted on 2019-03-18 18:46 UTC by Ms. Luz Barrero, Corporacion Colombiana de Investigacion Agropecuaria
RE: Welcome message - synbio forum - topic 5 [#9609]
Thank you Maria and Casper for your unyielding energy in guiding the topics of the forum.

I’d like to pick up on the suggestion that it is obvious that regulation must focus on the product of the techniques rather than on the process through which products are made. I’m afraid that this is not as obvious to me.

For example, techniques such as used in gene editing may, as some commentators have noted, result in changes as slight as single nucleotide substitutions or deletions. However, they also can be applied in series to create changes at a single locus almost to any scale of different, or be applied simultaneously to many loci to create something that in any pairwise comparison is ‘just like what might be found in nature’, but taken together is unlike anything found in nature.

In both cases, what is used as the comparator determines how a product is judged. In the first case, if the comparator is the previous edited product in the series, then the outcome of a ‘product’ view would be substantial equivalence. In the second case, if each change is taken separately, again the outcome might be ‘substantial equivalence’. Use a different comparator and the judgement could be very different.

Meanwhile, there is utility in process-informed regulation. For example, there was a recent case in the USA where plans for making plastic firearms using 3D printers were (at least temporarily because I don’t know the final outcome) banned from publication. The reasoning here is that waiting to regulate on the product, the plastic guns, was not the appropriate way to prevent the putative harm of plastic guns.

Moreover, where an LMO or a product of syn bio is not adequately able to be assessed using the comparative approach (eg, the discussion by Mr Durham [#9606]) for whatever reason, process can help to inform the approach to the risk assessment, such as in the search for hazards which is not informed by differences with a comparator.

Regards
Jack
posted on 2019-03-18 19:52 UTC by Mr. Jack Heinemann, University of Canterbury
RE: Welcome message - synbio forum - topic 5 [#9610]
Dear participants, thank you again for sharing views and submitting information. This week, under topic 5, we are discussing whether some organisms developed through new developments in synthetic biology may fall outside the LMO definition of the Cartagena protocol.

Clearly, there is no globally agreed definition on synthetic biology. Luz (#9608), Ben (#9606) and others have indicated that this causes some confusion. I agree with you, but at the same time, I think we have to live with that (at least for now). The picture is complicated. My view is that Parties would have a hard time negotiating and agreeing on a clear definition. Already at this stage, Parties regulate synthetic biology and genome edited organisms differently.

My advise is that we make use of the operational definition as provided by the Ad Hoc Technical Expert group on Synthetic Biology:

“Synthetic biology is a further development and new dimension of modern biotechnology that combines science, technology and engineering to facilitate and accelerate the understanding, design, redesign, manufacture and/or modification of genetic materials, living organisms and biological systems”.

Furthermore, that we keep in mind and respect that in 2016, Parties considered this definition "…useful as a starting point for the purpose of facilitating scientific and technical deliberations under the Convention and its Protocols." (Paragraph 4 of decision XIII/17 https://www.cbd.int/doc/decisions/cop-13/cop-13-dec-17-en.pdf ).

I agree with Ben (#9606) that the above mentioned synbio operational definition or "decription" creates an overlap between synthethic biology and traditional genetic modification. At the same time, I think we all share the view that synthetic biology potentially goes far beyond traditional genetic engineering, for instance by the design of genomes, artificial cells and/or introduction of entirely new engineered metabolic pathways encoded by extensive gene packages. This "depth of intervention" adds complexity.

The discussion under topic 5 is important. The Parties to the Convention has found it necesseray to address whether new synbio organisms are within the scope of the Cartagena Protocol (even though a synbio organism is not very well defined!). If we think that certain organisms are NOT COVERED by the Protocol, subsequently Parties may discuss ways to fill potential regulatory gaps in order to safeguard biodiversity. Furthermore, even for synbio organisms considered to be WITHIN the scope of the Cartagena Protocol, Parties may identify various challenges. For instance, it may be difficult to perform a risk assessment when a relevant comparator is missing and/or the organism expresses entirely new traits. These are all very interesting questions and topics.

However, our primary task now under topic 5 is to consider whether there are synbio organisms that do not fall within the scope of the Cartagena protocol.
Keep on posting!
Casper
posted on 2019-03-18 20:06 UTC by Mr. Casper Linnestad, Norway
RE: Welcome message - synbio forum - topic 5 [#9611]
Thank you Casper for this timely and helpful reminder on the working definition of synthetic biology adopted by the Parties.

“Synthetic biology is a further development and new dimension of modern biotechnology that combines science, technology and engineering to facilitate and accelerate the understanding, design, redesign, manufacture and/or modification of genetic materials, living organisms and biological systems”.

There are two aspects to my reply to the task of determining “whether there are synbio organisms that do not fall within the scope of the Cartagena protocol.”

First, there is the consideration of scope vs operational parameters of the Protocol. At present countries have little or no guidance for the risk assessment and risk management of LMOs that are not intended to be substantially equivalent, as we agree is a feature of anticipated synthetic biology products. The former AHTEG on RA&RM provided guidance firmly grounded in the comparative approach, noted that this would not always be adequate (eg, see Part II on abiotic stress), but then offered no general solutions to the problem. I would say that many products technically in scope are in urgent need of being made operational within the Protocol.

Second, the scope of the Protocol may be inappropriately limited depending on whether we approach the question from semantics or from risk. How much organic does something have to be to be an organism is a semantic approach. In contrast asking can a thing reproduce or act “to facilitate and accelerate the understanding, design, redesign, manufacture and/or modification of genetic materials, living organisms and biological systems” is a risk-based approach. As computing technology challenges the biological hegemony for possession of consciousness, so does it challenge the boundaries of organisms and biological systems. For example, is a robot with a DNA-based computer in the scope of the Protocol? Would it be if it could reproduce itself, or if it used some of its nucleic acid componentry to manufacture and/or modify genetic materials with or without the intention of doing so in other organisms?

The answer at this stage from my point of view is that the risk approach to determining the scope of the Protocol is the one better aligned with the Protocol’s objectives and spirit, and the precautionary principle.

Best to all
Jack
posted on 2019-03-18 20:34 UTC by Mr. Jack Heinemann, University of Canterbury
RE: Welcome message - synbio forum - topic 5 [#9615]
Dear Participants

I also concur with Casper on considering the definition of Synthetic Biology provided by  Ad Hoc Technical Expert group on Synthetic Biology. I still believe that  synthetic biology is an extension of genetic engineering techniques hence falls within the scope of  of the Cartagena Protocol. Lets maybe identify the gaps in Risk Assessment and Annex III to either expound or identify relevant clause/s in the protocol that will cover for all the gaps for either Synthetic Biology or any other related new techniques in future. As much as we are appreciating the new developments, lets not forget that protocol is our reference and focus on the gaps. for upcoming new developments.
posted on 2019-03-19 22:15 UTC by Ms. Motlalepula Pholo, Department of Agricultural Research
RE: Welcome message - synbio forum - topic 5 [#9617]
Dear Casper & colleagues,

I want to thank Maria, Casper and participants in this online forum for very interesting discussions.

With regard to the topic 5, I support the suggestion by Casper on considering the definition of Synthetic Biology provided by Ad Hoc Technical Expert group: “Synthetic biology is a further development and new dimension of modern biotechnology that combines science, technology and engineering to facilitate and accelerate the understanding, design, redesign, manufacture and/or modification of genetic materials, living organisms and biological systems”.

I do also want to highlight that synthetic biology would not been possible without the developments within genetic engineering. However, there may be some synthetic biology products that fall outside the Cartagena Protocols definitions of LMOs:  "...any living organism that possesses a novel combination of genetic material obtained through the use of modern biotechnology”. As for example organisms whose genomes has been edited without the use of nucleic acids using only proteins/protein complexes introduced into the cell. This has also been raised by Mr. Tzvetkov (#9605).

I consider the suggestion by Ms. Pholo (#9615) to identify the gaps in Risk Assessment and Annex III to be very relevant, and that such potential gaps may cause that the guidelines for risk assessment need to be revised or adapted. One option may be to look into how new developments within sequencing, metagenomics and proteomics can be used to fill such potential gap.

I would also like to highlight that the identification of a proper comparator (or isoline) (see also Mr Durham (#9606)) to be used in risk assessment may be a challenge for some products made by the “bottom up” approach of synthetic biology as  for example artificial cells.


Best regards from
Anne I. Myhr
posted on 2019-03-20 13:12 UTC by Ms. Anne Myhr, Norway
RE: Welcome message - synbio forum - topic 5 [#9638]
Thank you Casper for this refining of the question under discussion in this topic.

I agree with you and others that the operational definition of synthetic biology that we now use is really more of a ‘conceptual description’ than a definition and not really close to the precision and more prescriptive definition that we have for LMOs as written in the Cartagena Protocol.  Definitions generally ‘define’ things and our operational definition of synthetic biology has a lot of fuzziness around the edges. So this complicates our discussion of whether the organisms that result from the application of newer (synthetic) technologies are LMOs or not – nevertheless I am gratified to see the consensus gelling around the fact that all the examples we can think of are considered LMOs. This is really a tribute to the scientists and negotiators that crafted the LMO definition. It would appear to also simplify one of the tasks of the upcoming Ad Hoc Technical Expert Group meeting which must also answer this question – this will go a long way to refining the scope of which organisms we can talk about under this heading if we can also come to consensus on avoiding duplication.

Thanks
Jim Louter, Environment and Climate Change Canada
posted on 2019-03-22 18:55 UTC by Mr. Jim Louter, Canada
RE: Welcome message - synbio forum - topic 5 [#9614]
Thank you for the opportunity to contribute to this Forum. I am director of the Australian not-for-profit Gene Ethics, a citizens network. We want the precautionary principle, sound scientific evidence and robust laws rigorously applied to all proposed uses of genetic manipulation (GM) techniques and their products, including Synthetic Biology.

Though the Australian ACOLA report was first mentioned under Topic 1, I consider the following remarks still relevant in the present discussion of the scope of regulations under the Cartagena Biosafety Protocol. Note that the Australian government is not party to the Protocol, despite our many briefings refuting their spurious claim that the Protocol may be used to erect non-tariff barriers to trade.

Claudia Vickers [#9383] (CSIRO, University of Queensland, and co-author of the ACOLA horizon-scanning report) and Louisa Matthew [#9405] (Office of the Gene Technology Regulator Australia) both mentioned the ACOLA report, 'Synthetic Biology in Australia: an outlook to 2030', a 'horizon-scanning' study that the Office of the Chief Scientist of Australia commissioned. Neither updated this forum on the review of Australia's Gene Technology Regulations (as opposed to the Act) which has been ongoing since 2017.

Regulatory review

The ACOLA report (September 2018) asserts that the, "Australian regulatory scheme is based on a process trigger that captures all dealings with genetically modified organisms and focuses on the identification and management of risks to people and the environment. ... The primary legislation provides a broad definition of gene technology and GMO, and the regulations contain lists of excluded techniques and organisms based on an established history of safe use." p105

This account needs updating since Table 3 p106 (February 2018) shows the three reviews of the Gene Technology laws and regulations then underway.

1. By amending the Gene Technology Regulations, the Office of Gene Technology Regulator (OGTR) and the federal Health Department propose the deregulation of all SDN1 techniques (CRISPR, ZFN, Talen). This is the first occasion on which a whole class of techniques (and their living products) would be made exempt from any regulation at all, in an unspecified range of organisms. It would nullify the claim that the Gene Technology Act 2000 captures all dealings.

2. The deregulatory proposal also ignores the scant history of CRISPR's safe use in the genetic manipulation of humans, animals, plants and microbes. This renders false and meaningless the report's claim that techniques and organisms excluded in the Regulations must have a history of safe use. The contained and trial use of CRISPR SDN1 organisms is very limited and their commercial use is near zero. Yet new scientific evidence of off-target effects, mutations and various unexpected problems with CRISPR techniques is discounted and ignored. The advice on which the deregulatory proposals rest was from some experts with obvious commercial and scientific conflicts of interest that were undeclared and the ignored.

3. At the meeting of the Legislative and Governance Forum on Gene Technology on October 11 2018, state and territory ministers did not agree to the deregulatory proposal. Instead they sought further advice on the market implications of broad SDN1 deregulation, the area over which they exercise discretion. The advice has not been published. Our FoI requests rejected and Greens Senator Rice also requested the advice but this was declined.

However, at Senate Estimates hearings the head of the federal Agriculture Department stated in response to questions, "Our advice was that, while not without risk, the potential (export market) risks to adopting that (de)regulatory stance were not unmanageable. ... It  is  quite possible that for some markets the level of assurance and the investment in physical segregation systems may prove to be prohibitive. ... We weren't asked to do a comprehensive analysis. We were asked to provide some advice on the market access implications." http://emergingtech.foe.org.au/wp-content/uploads/2019/02/Rural-and-Regional-Affairs-and-Transport-Legislation-Committee_2019_02_19_6944.pdffileTypeapplication_pdf.pdf

Public engagement

The ACOLA report also says, "Hence policy makers, regulators and social scientists need to pro-actively engage the community and different interest groups to develop dialogue and build consensus on both benefits and risks and on the regulation of the field." p99.

However, the authors of the report fail to heed their own advice. During preparation of the report there were no processes of debate, discussion or information sharing with the general public or with us. When we heard the report was being prepared, our requests to the chair of the committee for a consultation received no response. We were not advised, for example, of a Fenner Conference on gene drives that joint sponsor of ACOLA, the Australian Academy of Science, held last November. Yet two presenters at the event knew of our interest and engagement with this use of the new GM techniques. https://www.science.org.au/news-and-events/events/fenner-conference-environment-use-gene-drive-technology-conservation

It appears that the establishment scientific and commercial sectors working on Synthetic Biology in Australia systematically exclude and marginalise both the general and interested publics. Such fortress and adversarial tactics will be counterproductive for us all.
posted on 2019-03-19 14:04 UTC by Mr. Bob Phelps, Gene Ethics (Australia)
RE: Welcome message - synbio forum - topic 5 [#9616]
Mr Phelps intervention (#9614) does not provide any information toward identifying examples of synthetic biology organisms that may fall outside the LMO definition of the Cartagena Protocol. Recognising it is outside the scope of topic 5, I will make my response brief.

As per Australian Government requirements on public consultation and transparency when developing regulatory measures, extensive information on the legislative review, including two calls for public submissions and the more than 1000 responses received, is available here: http://www.ogtr.gov.au/internet/ogtr/publishing.nsf/Content/reviewregulations-1. Application of gene editing does not necessarily result in synthetic biology organisms, and as such the review did not relate to synthetic biology. I reiterate my previously stated view on this (#9553), as shared by others on this forum (including #9536, #9588, #9435, #9446).
posted on 2019-03-19 23:58 UTC by Dr. Louisa Matthew, Office of the Gene Technology Regulator, Australia
RE: Welcome message - synbio forum - topic 5 [#9659]
Casper [#9610] has already noted that in this thread we are asked to focus on the topic of whether some organisms developed through new developments in synthetic biology may fall outside the LMO definition of the Cartagena. Notwithstanding, Mr Phelps’s intervention [#9614] requests engagement on the Australian regulatory environment and the Australian road-mapping report published by ACOLA. Louisa Matthews [#9616] has responded on behalf of the Australian Government’s Office of the Gene Technology Regulator regarding Australia’s regulatory environment. I will also make my response as brief as possible. As noted previously, I am a co-author on the ACOLA report. I have consulted with ACOLA in preparation of this response.

The synthetic biology report is part of a Horizon Scanning Series https://acola.org/programs/horizon-scanning-series/. The reports “draw on the deep disciplinary expertise from within Australia’s Learned Academies to analyse the future, navigate change and highlight opportunities for the nation. As interdisciplinary studies, ACOLA’s reports include economic, social, cultural and environmental perspectives to provide well-considered findings that inform complete policy responses to significant scientific and technological change.”

The ACOLA project team responded to all approaches by the community or individuals during the preparation of the report, including NGOs. ACOLA received literature from an NGO which was provided to the expert working group for consideration, and this was reviewed by the expert working group along with all of the other material. ACOLA reviewed their records but could not find any written correspondence from Mr Phelps when I enquired. The Chair has been contacted for comment and we are awaiting a response. Further enquiries may be directed to ACOLA.

The Australian Academy of Science is a member Academy of ACOLA. The member Academies of ACOLA hold their own activities and events and in this instance, ACOLA was not involved in the organisation or promotion of the Fenner Conference referred to. It was held after the launch of the Synthetic Biology report and as such ACOLA was not aware of it during the preparation of the report. As the Fenner Conference was run through the Australian Academy of Science, ACOLA was not involved in providing suggestions for potential speakers or participants/attendees.
posted on 2019-03-24 11:02 UTC by Dr Claudia Vickers, Commonwealth Scientific and Research Organisation
RE: Welcome message - synbio forum - topic 5 [#9619]
Dear Casper, dear all,

This is again an interesting and relevant topic for discussion.

My answer to this specific question would be the following.

A Living Modified Organism (LMO) is defined in the Cartagena Protocol on Biosafety as any living organism that possesses a novel combination of genetic material obtained through the use of modern biotechnology.
"Living organism" means any biological entity capable of transferring or replicating genetic material, including sterile organisms, viruses and viroids, and "Modern biotechnology" means the application of:
a. In vitro nucleic acid techniques, including recombinant deoxyribonucleic acid (DNA) and direct injection of nucleic acid into cells or organelles, or
b. Fusion of cells beyond the taxonomic family.

The operational definition of synthetic biology of the AHTEG synthetic biology is as follows: “synthetic biology is a further development and new dimension of modern biotechnology that combines science, technology and engineering to facilitate and accelerate the understanding, design, redesign, manufacture and/or modification of genetic materials, living organisms and biological systems”.

Based on these definitions, it can be considered that all living organisms developed through synthetic biology so far will fall under the definition of an LMO.  In the definition of an LMO,  the use of in vitro nucleic acid techniques  is not limited to only DNA as genetic material. Therefore living organisms that are built or modified to contain other genetic material than DNA (such as XNA) for replication would also fall under the definition of an LMO.
Examples of living organisms that fall outside the definition of LMOs seem therefore limited to living organisms that do not contain genetic material that can be transferred or replicated. This may apply to some living protocells that are still in an early state of research. 


Kind regards,
Boet
posted on 2019-03-21 13:17 UTC by Ms. Boet Glandorf, Netherlands
RE: Welcome message - synbio forum - topic 5 [#9621]
Dear participants,

Thanks again for the moderator and for the opportunity to comment.

Considering our task to consider whether there are synbio organisms that do not fall within the scope of the Cartagena protocol, in my opinion there is no example of a technological development that does not fit in the LMO definition and there is no concrete example of an aspect that would not be considered under the current risk assessment framework under the Cartagena Protocol or under existing regulations.

I would also like to support the #9616, and many others, regarding the fact that the application of gene editing does not necessarily result in synthetic biology organisms.

Best regards,
Luciana - Ministry of Agriculture
posted on 2019-03-21 14:09 UTC by Ms. Luciana Ambrozevicius, Brazil
RE: Welcome message - synbio forum - topic 5 [#9622]
This is my first post for some time so I will briefly introduce myself my name is Guy Reeves and I work at the Max Planck Institute for Evolutionary Biology in Germany. I have a background in molecular biology and evolution.

While it is often problematic to stretch the terms of the stated topic, I believe there is some value in considering the following :-

Consider whether any living intracellular organism/entity developed thus far through new developments in synthetic biology as per the Cartagena Protocol, might in some circumstances for compelling practical reasons, need to confer the status of living modified organism to its host (even when the hosts genome is entirely un-modified*)?

To take a current case example: using viruses genetically modified to express spinach defensin proteins in citrus trees. These viruses are not uniformly distributed in the tree and transgene expression is reported to be  generally stable for >7 years. The GM virus does not alter the tree chromosomes in any heritable* way .i.e the tree does not strictly meet the definition of and LMO (at least as I understand it).
While I will use this virus as a case study, much of what is written below would apply to any intracellular microorganism / or synthetic entity e.g. the “Virus-like macromolecular assemblies”  mentioned by Nikolay  [#9605]

Within  a single tree which is infected with the GM virus will be three types of tissues

a) tissues or fruits where no GM virus can detected.
b)  tissues or fruits where GM virus can be detected.
c)  tissues or fruits where it is conceivable the virus could be present but there is not data.

While it may be theoretically convenient to state that only the GM virus is of concern or regulated in practice as it is certainly a LMO. Is this a reasonable or useful position? As it is the movement or trade of tissues or fruits which is the only plausible focus for informed regulation (see below, point 2 of Ben”s post [#9606]).

While, outwardly this question is related to the unresolved debate about the grafting of GM plant root stocks onto non-GM plants,  the question is substantially modified by the fact that microorganisms have evolved to be infectious both from cell-cell but also between plants (in the above study plant to plant transmission is reported to be “unlikely” see .pdf) and “virus-like macromolecular assemblies” will presumably be designed to have similar properties.

Might the hosts of a LMO (e.g of a GM  virus, GM endosymbiotic bacteria, Virus-like macromolecular assemblies [#9606] ), in practice need to be considered as a living modified organism? Put another way, is stating that a potentially hard to detect microorganism is all that needs to be considered a responsible or practical position?  The importance in answering  this question, will presumably be heightened where microorganisms are readily  transmissible in the environment and or highly persistent in the environment.

Referencing the  very relevant exchange on how to consider substantial equivalence /substantial change in impact/ the appropriate comparator, for Synbio products between Ben [#9606] and Jack  [#9609], [#9611], Anne [#9617] I think it is notable that extensive regulatory process by both the EPA and USDA has, at least in my reading,  placed the test of substantial equivalence on both the GM virus AND the fruit.  With considerably more emphasis on the fruit. This can be interpreted to be an indirect acknowledgement that the host (which does not fit the definition of LMO) and not the actual LMO is the most practical  focus for regulation / discussion. This is why the definition  of LMO as per the Cartagena Protocol might need to be reconsidered in these set of up-coming circumstances.

This question goes far beyond the details of the case study I used to frame this question- in my opinion. Particularly, if as per Ben’s useful recommendation [#9606] we aim to address “2. We are aiming (ultimately) to provide regulatory guidelines -as already provided for LMOs- for Synbio.”


I am not considering epigenetic or transient expression changes.

References and additional details of case study in Citrus_GM_viruses_CBD.pdf
posted on 2019-03-21 15:56 UTC by Dr. Guy Reeves, Max-Planck-Institut für Evolutionsbiologie
RE: Welcome message - synbio forum - topic 5 [#9623]
Dear colleagues,

once again I would like to thank Maria and Casper for moderating this round of discussion.

As to the question at hand under topic 5 I would like to provide the following brief contribution:

A possible case may be living organisms generated by multiplexed DNA free ZFN/TALEN/MN applications, which can be considered SynBio. Such systems may be targeted to modify the genotype (through SDN-applications) or the phenotype by modification of gene expression through epigenetic mechanisms (through SDNs modified with domains that impact regulation of gene expression, repressors, positive transcription factors, or domains to introduce epigenetic modifications, e.g. methylases, demethylases, etc.)  (see e.g. Tycko 2017).

Tycko, J., Hess, G.T., Jeng, E.E., Dubreuil, M., and Bassik, M.C. (2017). The expanding CRISPR toolbox [Online]. Nature Methods. Available: http://s3-service-broker-live-19ea8b98-4d41-4cb4-be4c-d68f4963b7dd.s3.amazonaws.com/uploads/ckeditor/attachments/7742/CRISPR_poster-WEB.pdf [Accessed 27.11.2018].

Thank you and I am looking forward to our further debate. Kind regards

Helmut Gaugitsch
Environment Agency Austria
posted on 2019-03-21 16:29 UTC by Mr. Helmut Gaugitsch, Austria
RE: Welcome message - synbio forum - topic 5 [#9624]
Dear Colleagues,

This is almost verbatim of a post of mine under Topic 3 of the previous online forum, which unfortunately I can’t find. Apologies if I am repeating myself, but it might still be of some use, especially in the light of the comments of Guy (#9622) and Helmut (#9623).

The following are just some possible cases where further analysis might be needed to decide if the organisms should or should not be considered LMOs as defined in the Protocol.

-> 1. The entity in question is a living organism that possesses a novel combination of genetic material, but is NOT obtained through the use of modern biotechnology
The discussion on gene editing falls under this category and is probably the trickiest of all examples provided. Here, let me please try to deconstruct the process of creating an organism (i.e. gene editing), which is the part that is relevant to our discussion, from the discussion on whether or not biosafety measures (e.g. risk assessment, labelling, etc) should be imposed under such organisms.
The techniques for gene editing, such as CRISPR, Meganucleases, Zinc-finger nucleases (ZFN), Transcription activator like effector nucleases (TALEN) or oligonucleotide directed mutagenesis, all use nucleic acid techniques to trigger modifications in genomes.
Gene editing normally requires the introduction of new genetic material (expression construct) into the cell. The organism thus generated will be an LMO. But that expression construct can be removed from the cell after the modification is accomplished. In some of those cases when only single point mutations, deletions or small insertions are introduced and if the recombinant construct is no longer present it may not be possible to distinguish whether a modified organism was produced by use of classic mutagenesis or by modern biotechnology.
In addition some of the modifying agents used for gene editing, such as site-specific nucleases or oligonucleotides, can be introduced into the cell directly for example with the help of liposomes or electroporation. Then the resulting modified organism might fall outside of the scope of the definition of an LMO as it was generated with the help of a sophisticated mutagenic reagent.
When we discuss gene editing vis-à-vis the definition of modern biotechnology (“In vitro nucleic acid techniques … that overcome natural physiological reproductive or recombination barriers …”), I trust that we may all agree that the different methodologies all involve a “nucleic acid” technique in one step or another of the process. However, I can foresee some controversy on whether or not the techniques “overcome natural physiological reproductive or recombination barriers”. As such, it might be hard or practically impossible, with the current definition of an LMO under the CP, to determine whether or not an organism is an LMO from a legal point of view.

-> 2. The entity in question is a living organism that possesses genetic material from the same species or sexually compatible species
It is possible to introduce changes in that already occur in the same species or in closely related sexually compatible species, through a process usually known as cisgenesis. If those are the only modifications introduced the resulting organism may be considered equivalent to an organism that has been produced by traditional breeding or selection. In such cases the use of modern biotechnology would not be considered as a means to “overcome natural physiological reproductive or recombination barriers”.

-> 3. The entity in question is a living organism that possesses a novel combination of genetic material obtained through the use of modern biotechnology, but that genetic material has no similarity with any naturally occurring genetic material
I am personally well aware of an example of that kind. The laboratory of Dr Michael Hecht designed de novo proteins on the basis of simple amino acid pattern that share no similarity to any natural proteins and whose only significant physicochemical property is to form β-amyloid (DOI: 10.1073/pnas.96.20.11211). It was shown by Dr Ulrich Hartl, Dr Martin Vabulas and their colleagues that when produced inside the cell those proteins can have profound effects on cellular processes (DOI: 10.1016/j.cell.2010.11.050).
In such cases it would be considered that any organism carrying such novel genetic material would be an LMO, because they fulfil all conditions in the definition of the Cartagena Protocol ,including the fact that natural physiological reproductive or recombination barriers would have to be overcome as they include limits to which random genetic material can be introduced into the genome.

-> 4. The entity in question is a living organism that has been synthesised entirely in vitro
If the whole genome has been synthesised entirely in vitro (possible for viruses and some bacteria, expected to be possible for higher eukaryotes soon) we have in vitro use of nucleic acid techniques. So if it possesses novel combination of genetic material it would be an LMO, if it is entirely identical to naturally occurring genome it may not be considered an LMO.

-> 5. The entity in question is a living organism that possesses a novel combination of genetic material, but only in cells and tissues that do not participate in sexual and non-sexual reproduction under natural conditions
It is possible to generate an organism where only some of the cells or tissues are modified, e.g. by grafting. Those modified tissues or cells might not be able to participate in sexual (as germ cells) or asexual reproduction under the natural conditions for the species in question nor to exchange genetic material with other cells that can participate in such reproduction. The second point is not trivial because it was shown in the laboratory of Dr Ralf Bock grafting can result in exchange of genetic material (DOI: 10.1038/nature13291) and chloroplast genomes can be transferred horizontally as well (DOI: 10.1073/pnas.1114076109). The status of such organism as an LMO is not very clear, because it won’t be able to transfer the modified genetic material, but it still might be able to replicate it even when it is sterile. Therefore, only in situations where the cells carrying the novel combination of genetic material are not capable of transferring or replicating would the organism not be considered an LMO.

-> 6. The entity in question in not living organism
For examples under this category to be excluded from the LMO definition, the entity in question must not be capable of transferring or replicating genetic material. One such example may be prions, which does not contain genetic material as such and are not specifically included in the definition of living organism. Another one might be those virus-like assemblies mentioned above.

I hope that this rather long repletion of almost two years old view might still be of some help to clarify the topics being discussed.
Best Regards,
Nikolay
posted on 2019-03-21 17:00 UTC by Mr. Nikolay Tzvetkov, Bulgaria
RE: Welcome message - synbio forum - topic 5 [#9625]
Dear colleagues

I would like to mark my appreciation for how thoughtful and constructive this forum has been.

A few points in summary are for me worth highlighting. (1) The invention of new deoxyribonucleic acid bases, as mentioned most recently in post [#9616] (Boet), makes clear that LMOs can be made using genetic material other than the four currently used bases of DNA. I am glad for this recognition given how long RNA as been sidelined from thorough consideration as a way to genetically modify organisms. RNA has the same capacity to carry genes, does so in some kinds of viruses, perpetually replicates in some kinds of organisms (e.g. yeast) as RNA, and has well known ways to convert back and forth from DNA and be used to make proteins. Perhaps the recognition of xeno-bases will elevate RNA based technologies to their uncontested place of coverage under the Protocol if only as another form of xeno-nucleic acid, where xeno means other nucleotides.

(2) The highly literal examination of the words in definitions included in the text is an important anchor for these discussions, but it is also potentially limited in its value for determining the relevance of techniques and products that were or are not now envisioned by current understanding of what those words meant then. The use of the term ‘nucleic acid’ can be seen in context to mean material that can be the basis for inheritance because that was the dominant impression of what genes were at that time. Constraining ourselves now to the literal polymer view is not the equivalent of the modern spirit of the Protocol (in my opinion). Moreover, this approach shifts the debate to other, undefined, terms. What is ‘living’? Is the virus-like macromolecular assembly (ie robot in my terminology) described in [#9605] (Nikolay) not composed of a nucleic acid that is ‘injected’ into an organism? How would you ever absolutely prevent its recombination or replication completely assuming that was even the intention? See [#9622] (Guy) for a thoughtful discussion on this.

Therefore, it might be found that such things pose little or no (unmanageable) risk, but defining them outside of the Protocol is to make an a priori decision that if no risk assessment was done that would be okay. That seems to use a technical rationale based on the current understanding of what words meant when the Protocol was written, rather than what the words were possibly intended to mean.

(3) Unlike others, I would extend this discussion to so-called “transient” or “epigenetic” changes, for two reasons. First, the Protocol talks about modification, not the number of generations for which that modification persists. Generation time is a poor metric because it has such an enormous biological range. A virus generation time is the number of years that it can be reactivated from a long-lived host or the shortest time to replication in a newly infected host? (Does a modification that lasts only 2 seasons on organisms purposely spread over a million hectares have no risk while a modification that potentially lasts ‘forever’ in the germline of something in the laboratory worthy of a risk assessment?) To pivot on ‘transience’ is to again argue about what that word means and is a problem I think that only arose post-writing of the Protocol. Second, changes to molecules that are involved in gene expression are not epigenetic changes, despite the increasingly common conflation of gene expression with epigenetics. We don’t need to call the mechanisms of turning genes on and off epigenetic unless we mean that they are indeed heritable, which is where the genetic part of the word epigenetics becomes meaningful. So if such things are heritable modifications, they create the potential for an organism to have all or some of the risks that LMOs created through DNA changes can have.

Thank you again colleagues for your patience with such long posts and the vibrant discussion.
Jack
posted on 2019-03-21 19:45 UTC by Mr. Jack Heinemann, University of Canterbury
RE: Welcome message - synbio forum - topic 5 [#9627]
Dear Casper, dear all,

Acknowledging the risk of having a slightly circular discussion, I would like to pick up a few points from Nikolay, Jack, Anne Myhr, and also Casper.

Starting with Jack’s second contribution [#9611], I liked his approach to the interpretation of the scope of the protocol, i.e. that we can tackle the issue either through the lense of semantics or that of risks - “the scope of the Protocol may be inappropriately limited depending on whether we approach the question from semantics or from risk.”

If we take the example of an organism whose nucleic acid has been intentionally modified at a predetermined location in the lab by a tool of modern biotechnology. If the tool is a pre-designed protein (or a protein complex) -the example chosen my Nikolay [#9605] and Anne Myhr [#9617]-, an endonuclease such as ZFNs or Talens,  why would it matter if the protein was produced outside the organism in a testtube or flask  instead of being produced within the organism after supplying the organism with the DNA instructions for producing that protein (either transiently or integreted as a transgene into the genome)? In both cases it’s a nucleic acid technique, as it is the nucleic acid of that organism that is being altered in a very specific and predesigned way, through the use of modern biotechnology. I actually do not want to go down the road of semantics here, but rather stay with the task of what is important. Is it about using the CPB as widely and as openly as possible to ensure safety in line with the precautionary approach? – or do we want to interpret it as narrow as possible and battle over each and every word in order to get as many LMOs excluded as possible?  This is a very interesting question, well worth while pondering over.

If we agree on a wide scope and a broad interpretation then we will probably have all organisms currently produced through synthetic biology covered by the CPB. If not, then we need to clarify and ensure that they will be covered in other ways under the CBD to ensure environmental and human safety, including socio-economic considerations.

It is this, that Casper reminded us to be our task, namely to identify if -so far- all organisms produced through synthetic biology are LMOs covered by the Protocol and if not, which ones are not covered. I will follow that request and refrain from following the temptation to go back to the tricky issues of suggested LMO excemptions. (will keep those thoughts separarte -at least for now).

Finally, thank you Jack for thinking outside of the box. We need that! Like what is there that is using nucleic acid (whether DNA, RNA, XNA) or has it an insturctural component that is different from the normal LMO?

I will keep thinking, but haven’t quite got there yet.

With kind regards,

Ricarda
posted on 2019-03-21 23:27 UTC by Ms. Ricarda Steinbrecher, Federation of German Scientists (Vereinigung Deutscher Wissenschaftler)
RE: Welcome message - synbio forum - topic 5 [#9628]
Dear all—

My thanks to our moderator and to all for a very interesting discussion.

I would like to simplify Nikolay’s excellent (but complex) posting [#9624] into just two ends of a continuum: 1) are there organisms developed using synthetic biology that are so different that they fall outside the definition of LMO under the Cartagena Protocol and 2) are there organisms so similar to those found in nature that they fall outside the Cartagena definition of LMO from the other end?

Starting with the first end of the continuum, I agree with Boet [#9619] and Nikolay: nothing so different that it falls outside the Cartagena definition of LMO. 
To my knowledge, there is still but one example of a reproducing organism with an expanded genetic alphabet (4 DNA bases plus 2 additional XNA bases), which its developers call a “semisynthetic organism”.  It is an LMO as defined by the Cartagena Protocol. http://www.pnas.org/content/114/6/1317.full?sid=8fd587ac-be94-40c0-b27f-f3320986ad6c.

Boet also mentions an active area of research to develop bottom-up, artificial cells called “protocells”.   I know of no example of a self-replicating protocell today.  In addition, most of the research groups working in this area are exploring protocells based on nucleic acids, which if successful, would also be considered an LMO under the Cartagena Protocol.

At the other side of the continuum, are there organisms developed using synthetic biology methods that are so similar to those found in nature that they fall outside the Cartagena definition of LMO from the other end?  Louisa [#9616], Luciana [#9621] and quite a few participants during the earlier topics point to examples of genome editing where they believe this is the case.  LMO’s must possess “a *novel* combination of genetic material obtained through the use of modern biotechnology”.  And as Nikolay points out, “modern biotechnology” means the application of techniques “that *overcome natural* physiological reproductive or recombination barriers”.  (asterisks added for emphasis.)  Indeed, in my view, one of the significant “new dimensions” of synthetic biology is that its approach to “understanding, design, redesign, manufacture and/or modification of genetic materials and living organisms” (following the AHTEG operational definition) now enables scientists and engineers to design organisms that are virtually indistinguishable from those that might occur through traditional breeding.

Best regards,
Bob Friedman, JCVI
posted on 2019-03-22 03:04 UTC by Mr. Robert Friedman, J. Craig Venter Institute
RE: Welcome message - synbio forum - topic 5 [#9629]
Dear All,

I totally agree that organisms derived thought synthetic biology at current stage fall in the scope of LMOs of the protocol. However, there might be some gaps in risk assessment, e.g.   appropriate non-modified comparator, as mentioned in the past online forum discussion on RA &RM (UNEP/CBD/BS/RARM/AHTEG/2016/1/5). In addition, as genome editing employs or results in nucleic acid recombination, organisms derived from this technology shall also fall in the scope of the protocol.

Wei
posted on 2019-03-22 03:54 UTC by Mr. Wei Wei, China
RE: Welcome message - synbio forum - topic 5 [#9631]
Dear colleagues,

Thank you very much to all of you who already provided some input to the question whether any living organism developed through synthetic biology fall outside of the LMO definition.

I would like to join those (e.g. #9619, #9621, #9628, #9629) who stated that so far there are no living organisms that fall outside of this definition. I also agree with post # 9619 that living organisms with genetic material other than DNA also fall under the definition of an LMO.

Finally, I would like to reiterate that genome editing per se is not synthetic biology. Genome editing is a set of techniques that can result in changes going from a single nucleotide to whole genes/genomes and is used in a range of applications in modern biotechnology and synthetic biology. This has been stated by many delegates at COP14 and resulted in decision 14/19, Terms of reference for the AHTEG, which asks the AHTEG to take stock of “concrete applications of genome editing if they relate to synthetic biology”. 

Best regards,
Swantje
posted on 2019-03-22 11:44 UTC by Ms. Swantje Strassheim, Germany
RE: Welcome message - synbio forum - topic 5 [#9632]
Hello again, dear participants.  To follow along with what #9553, #9621, and others have shared about the degree to which some products of synthetic biology can be considered "novel," I'd like to share that in the U.S. experience, when making a policy decision regarding the safe handling and use of a modified organism, it is most valuable to consider: 1) the characteristics of the organism in question, and 2) the novelty of those characteristics.  There are numerous internationally recognized resources in existence that provide frameworks for how to consider the characteristics of organisms:

United States Environmental Protection Agency (EPA):
https://www.epa.gov/risk/risk-assessment-guidelines

International Plant Protection Convention (IPPC):
https://www.ippc.int/en/core-activities/capacity-development/training-material-pest-risk-analysis-based-ippc-standards/
http://www.fao.org/docrep/009/a0450e/a0450e00.htm
http://www.acfs.go.th/sps/downloads/34163_ISPM_11_E.pdf

Organization for Economic and Cooperation and Development (OECD):
http://www.oecd.org/chemicalsafety/biotrack/oecdandrisksafetyassessmentinmodernbiotechnology.htm 


World Organization for Animal Health (OIE):
http://www.oie.int/en/our-scientific-expertise/specific-information-and-recommendations/invasive-alien-animal-species/

World Health Organization (WHO):
http://www.who.int/tdr/publications/year/2014/guide-fmrk-gm-mosquit/en/

Many thanks to all for this constructive dialogue!
Best - Jenna
posted on 2019-03-22 12:33 UTC by Ms. Jennifer Shinen, U.S. Department of State
RE: Welcome message - synbio forum - topic 5 [#9633]
Dear all,
I would like to support the statements made by Swantje Strassheim #9631 and by Jennifer Shinen #9632 regarding the fact that assessments need to be made on a case by case basis and that so far there are no living organisms that fall outside of the LMO definition. This also applies for organisms containing a gene drive mechanism.
I agree with post #9632 listing existing framework that are available to assess those LMOs.
Thank you all for the great dialogue.
Best
Delphine
posted on 2019-03-22 13:20 UTC by Ms. Delphine Thizy, Imperial College London
RE: Welcome message - synbio forum - topic 5 [#9635]
I also support the opinion that those synthetic biology organisms developed so far fall inside the definition of LMO. However, this might change quickly and it is, therefore, of paramount importance to follow the progress in this field closely. As Swantje Strassheim pointed out  [#9631], it is not the technology that defines product to be or not to be a product of syntetic biology, but the specific application of genetic engineering. Thus, I do not see any alternative to an assessment on a case by case basis, as Delphine Thizy  [#9633] and others pointed out.
Regards
Christoph
posted on 2019-03-22 16:16 UTC by Mr. Christoph Lüthi, Switzerland
RE: Welcome message - synbio forum - topic 5 [#9637]
It is a pleasure to contribute to the debate of Topic 5, aiming to address if synthetic biology organisms may fall outside the definition of living modified organisms as per the Cartagena Protocol. This is not a new issue considering the synthetic biology debate in the past years.

In my opinion, synthetic biology organisms and products are within the framework of the LMOs definition. The online and ATHEG meetings in 2015 (2015 AHTEG report: UNEP/CBD/SYNBIO/AHTEG/2015/1/3) and 2017 (2017 AHTEG report: UNEP/CBD/SYNBIO/AHTEG/2017/1/3) already discussed this question.

Considering the doubts and the proposal to get back to this agenda due to the debates around new developments in synthetic biology, it is quite relevant to stress that living organisms already developed or under research and development using synthetic biology techniques are LMOs as defined by the Cartagena Protocol.

Synthetic biology is comprised by the scope of biotechnology, given the definition of the CBD, and by modern biotechnology as the definition of the Cartagena Protocol. It is reasonable to argue that all the examples of synthetic biology discussed in the past years an in the present online forum are LMOs as defined by the Cartagena Protocol.

The only exception are mutants developed using genome editing, that are developed based on conventional methods and cannot be, by definition, considered LMOs.

Topic 5 is extremely important considering the role of risk assessment under the Cartagena Protocol and the related experience of LMOs.
posted on 2019-03-22 18:30 UTC by Mr. Rodrigo C A Lima, Agroicone Ltd
RE: Welcome message - synbio forum - topic 5 [#9639]
Dear Colleagues, dear Casper and Maria,
this is really an interesting discussion. I would like to answer to post # 9606. Thank you Ben David, for summing up the two purposes. But in my opinion the most important purpose/question is missing. The most important question is:
Can organisms produced through Synthetic Biology adversely affect the three objectives of the convention, so that the scope of the Cartagena-protocol does not address the risk?
Or in different words: Are there any living organisms produced through Synthetic Biology (in its broad definition) that, if they do not fall under the scope of the Cartagena-Protocol, e.g. because they are not transboundary moved, can adversely affect biodiversity?
The most important target of the horizon scanning is, to be aware of developments of synthetic biology that may have negative effects on the biodiversity. For that aim, it is relatively unimportant, if they fall under the definition of LMOs of the Cartagena-Protocol.
I would also like to answer to your post #9610 Casper: is the question only, if we think that certain organisms are not covered by the Protocol, or is it also that the area of the application of a certain organism is not covered by the Protocol?
Plus as I already mentioned in my post to topic 1. It is not only the organism we have to consider, but also the way it is used. Therefore, I would like to highlight that as #9615 says, synthetic biology is an extension of genetic engineering. Which for my understanding means, it is more and it goes far beyond. Therefore, we need to consider everything. Not only if the organism is still an LMO but also how far does its use go, how far does it affect nature, what is the aim of its application, etc. 
Also in reply to Ricardas Post #9627: only because organism are defined as LMO, there use is not always covered by the scope of the CP as the objectives of the CP do not address all objectives of the CBD.
So I hope that my post does not exceed the scope of topic 5 to much,
Birgit
posted on 2019-03-22 20:19 UTC by Ms. Birgit Winkel, Germany
RE: Welcome message - synbio forum - topic 5 [#9640]
Dear Participants,

In consideration of the topic raised for this topic, research activities framed in synthetic biology are currently carried out that would be beyond the scope of the definition of modified living organisms of the Cartagena Protocol.

To cite some approximations of what might be beyond the scope of the Cartagena Protocol, we found a study developed in 2016 by researchers from the Craig Venter Institute, the Microscopy and Images Research Center of the University of San Diego, the Synthetic Genomics company, and the Institute of Standards and Technology in the United States, who used the complete genome design and complete chemical synthesis to minimize the synthetic genome of 1079 kilobases of Mycoplasma mycoides JCVI-syn1.0. After several attempts at design, synthesis and testing, with retention of almost essential genes, they were able to obtain JCVI-syn3.0, with 531 pairs of kilobases and 473 genes, this being a smaller genome than that of any cell with autonomous replication. is in nature. JCVI-syn3.0 retains almost all the genes involved in the synthesis and processing of macromolecule, as well as 149 genes with unknown biological functions. According to the authors, the creation of JCVI-syn3.0 can be considered as a versatile platform to investigate the basic functions of life and to explore the design of the whole genome (Hutchison III, et al., 2016).

Another even more recent publication, carried out by researchers from the Engineering Research Center of Synthetic Biology in China, shows the proposal of what they call Project Sc2.0, by means of which the construction of a yeast genome is proposed to demonstrate the ability to build a yeast with multiple synthetic chromosomes and at the same time allow to answer important biological questions (Dai, et al., 2017).
 
Hutchison III, C. Chuang, R-Y, Noskov, V., Assad-Garcia, N., Deerinck, T.J. Ellisman, M.H., Gill, J., Kannan, K., Karas, B.J., Ma, L., Pelletier, J.F, Qi, Z-Q., Richter, R.A., Strychalski, E.A. Sun, L., Suzuki, Y., Tsvetanova, B., Wise, K.S. Smith, H.O., Glass, J. Merryman, C., Gibson, D. & Venter, C.; 2016; Design and synthesis of a minimal bacterial genome; Sience; 351(6280):6253-1 - 6253-11.
posted on 2019-03-22 21:57 UTC by Mr. Carlos Augusto Ospina Bravo, Colombia
RE: Welcome message - synbio forum - topic 5 [#9641]
Dear participants, again, thank you for sharing views and contributing to a fruitful discussion! Responding to Birgit's question (#9639) just now on whether we are to discuss also "the area of the application of a certain organism….", I think that under topic 5, our mandate is more narrow. We are only to discuss whether or not an organism developed through techniques of synthetic biology may fall outside the DEFINITIUON of living modified organisms as per the Cartagena Protocol. As I see it, we are not going into issues like transboundary movements, transit or various uses or applications. Birgit is raising a very valid point, though, in that the "trigger" for the Cartagena Protocol is a transboundary movement of a LMO. But, for now, we are to focus on the LMO definition and synbio organisms, and not increase the complexity!
Best wishes,
Casper.
posted on 2019-03-22 21:58 UTC by Mr. Casper Linnestad, Norway
RE: Welcome message - synbio forum - topic 5 [#9642]
Dear all,

Bob Friedman again, from the J. Craig Venter Institute (JCVI).  I would like to briefly respond to Carlos’ recent posting [#9640] that refers to research organisms developed by scientists at JCVI, Mycoplasma mycoides JCVI-syn1.0 and Mycoplasma mycoides JCVI-syn3.0.  These organisms, constructed using the techniques of synthetic biology, are indeed LMOs under the definition of the Cartagena Protocol.

As Carlos stated, the genome of syn 3.0, which took several years to design, is smaller than any autonomously replicating cell found in nature. http://science.sciencemag.org/content/351/6280/aad6253.  For simplicity, we called this a “minimal cell”, because if we removed any of its remaining genes, the bacterium would no longer be able to live and reproduce.  The goal of this research is to better understand the minimal requirements for life. 

Likewise, the international Sc2.0 Project Consortium is synthesizing a custom-designed genome for yeast, to serve as a platform for systematic studies of eukaryotic chromosomes.   http://science.sciencemag.org/content/355/6329/1040 .  Some, but not all, of the 16 chromosomes of the yeast Saccharomyces cerevisiae have been redesigned, to date.  This organism, constructed using the techniques of synthetic biology, is also an LMO under the definition of the Cartagena Protocol.

Regards,
Bob Friedman
posted on 2019-03-22 23:26 UTC by Mr. Robert Friedman, J. Craig Venter Institute
RE: Welcome message - synbio forum - topic 5 [#9644]
Dear Casper, dear all
It is a pleasure to contribute to the debate on Topic 5

I believe that all living organisms already developed or under research and development using synthetic biology techniques are LMOs as defined by the Cartagena Protocol.

Yet it is important to highlight that under synbio we are not just talking about living organisms but mainly parts and products of living organisms that result from Synbio and these are not well covered under the Cartagena Protocol. Also, when it comes to Synbio we need to keep in mind the issue of digital sequence information and the effect on the third objective of the CBD

Best regards,
posted on 2019-03-23 09:36 UTC by Mr. Ossama Abdelkawy, Syrian Arab Republic
RE: Welcome message - synbio forum - topic 5 [#9645]
Thank you dear moderator and dear participants for your interesting discussions

I think the 2015 AHTEG definition on synthetic biology is made perfect fit to synbio for understanding biological processes through their reconstruction and /or modification of genetic materials, organisms and biological systems, and the works that have been being done so far belongs inside this sympathetic and/or the definition of living modified organisms as per the Cartagena Protocol. Therefore, the concern of the discussion shall be how to regulate the new functionalities that arise from the modification. However, if there are synbio which deals with construction of new biological processes with new functionalities that we haven’t and can’t imagine, the AHTEG shall open a separate discussion/title.
posted on 2019-03-23 11:08 UTC by Mr. Taye Birhanu, Ethiopia
RE: Welcome message - synbio forum - topic 5 [#9648]
Dear colleagues,

Thank you once again Casper for moderating this important topic. And thanks to many colleagues who have brought very insightful comments to this discussion.

I would like to share some of the thoughts that have been the basis of my opinion that all organisms already developed or under research and development using synthetic biology related techniques are (and should be) LMOs as defined by the Cartagena Protocol.

The determination if an organism falls within or outside a regulatory ‘definition’ requires an interpretation of the law. This is an intrinsically difficult task, and as with any kind of text interpretation, it might be influenced by the values of the judge. Even lawyers and law makers, who have been trained to be unbiased, they can diverge and that is mainly the reason why most countries have collegiate court decisions.

I struggle with the need to have a strong and objective operational definition and the need to consider the ‘definition’ beyond its, perhaps broad or weak, wording. If in one hand focusing in the wordings makes the implementation easier, on the other hand, the Convention and Protocol objectives should be prioritized. Therefore, I tend to agree with other colleagues who have highlighted the need to interpret the definition beyond a literal examination of its wording and within the scope and objectives of the Protocol when it was first conceived. In fact, the is the way the European Court of Justice has interpreted the European GMO Law in the case of organisms obtained by means of techniques/methods of mutagenesis:

“[…] cannot be interpreted as excluding, from the scope of the directive, organisms obtained by means of new techniques/methods of mutagenesis which have appeared or have been mostly developed since Directive 2001/18 was adopted. Such an interpretation would fail to have regard to the intention of the EU legislature, reflected in recital 17 of the directive, to exclude from the scope of the directive only organisms obtained by means of techniques/methods which have conventionally been used in a number of applications and have a long safety record. It follows that an interpretation of the exemption in Article 3(1) of Directive 2001/18, read in conjunction with point 1 of Annex I B thereto, which excludes organisms obtained by means of techniques/methods of mutagenesis from the scope of that directive, without any distinctions, would compromise the objective of protection pursued by the directive and would fail to respect the precautionary principle which it seeks to implement.”

The Explanatory Guide to the CP also presents the need to consider these techniques in a wider context, which does not require the insertion of genetic material (Box 15, IUCN 2003). Basically, the idea was to separate techniques with history of safe use and novel techniques without history of safe use; the same interpretation of the latest ECJ.

Thank you,

Sarah
posted on 2019-03-23 12:54 UTC by Ms. Sarah Agapito-Tenfen, Brazil
RE: Welcome message - synbio forum - topic 5 [#9649]
Dear All,
Let me first thank Casper and Maria for moderating this complex and interesting online forum. And thank you all for the stimulating contributions.
I would like to start, as Casper suggested, from the definition of LMO under the Cartagena Protocol. This definition refers to “a novel combination of genetic material obtained through the use of modern biotechnology” and in this regard I think it is sufficiently broad to cover synthetic biology applications.
I agree with Casper (#9610) on the advice to use the operational definition of synthetic biology as proposed by the AHTEG on synthetic biology in 2015. I believe it is a good description of the domain, clearly indicating that synthetic biology is a further development of modern biotechnology. While showing new and broader potential, synthetic biology is the result of technological and scientific developments in biotechnology and other sectors which evolve in a continuous manner. The Scientific Committee on Consumer Safety (SCCS), the Scientific Committee on Health and Environmental Risks (SCHER) and the Scientific Committee on Emerging and Newly Identified Health Risks (SCENIHR) in 2014 concluded that it is difficult to identify criteria able to unambiguously differentiate synthetic biology from genetic modification*.
On this basis, I share the view of Motlalepula (#9615), Boet (#9619) and others that organisms developed through synthetic biology, currently and in the foreseeable future, fall under the definition of LMO in the CP. This was also the conclusion of the experts in the AHTEG and in the Scientific Committees mentioned above.
I read previous posts with great interest and I would like to try to limit the complexity of this exercise. Some of the comments provided in the discussion are very important but they seem to me more related to the interpretation of the LMO definition of the CP or to the scope of the CP rather than to real challenges arising from synthetic biology applications: e.g. whether gene editing can be considered modern biotechnology as per the CP definition (Nikolay, #9624) or whether the use of certain LMOs is not covered by the CP (Birgit, #9639) are important considerations, but they are not specific for synbio, they might apply to organisms other than those produced through synthetic biology.
I think that for the present objective we should be focused, our aim is to remain updated on the ongoing developments in synthetic biology and monitor whether current governance systems are adequate to ensure the three objectives of the Convention. In this respect the horizon scanning process on Synbio, this online forum and the submission by Parties are already providing useful contributions.
*SCENIHR, SCCS, SCHER (2014) Synthetic Biology I Definition, Opinion, September 2014. Available from: http://ec.europa.eu/health/scientific_committees/emerging/docs/scenihr_o_044.pdf
posted on 2019-03-23 16:26 UTC by Ms. Ilaria Ciabatti, European Commission
RE: Welcome message - synbio forum - topic 5 [#9655]
I express my thanks to forum participants for the interesting exchange of ideas. Many ideas that have been shared here are natural extensions to topic 5 and go to broader issues, but I agree with Ilaria [#9649] that we need to stay focused in our discussion and keep it specific to synthetic biology. For topic 5, this is living organisms (ie not components or products) developed thus far (ie what is known now, rather than what may be possible in future).

In an earlier post Robert [#9628] provided an elegant presentation of a challenge he perceives in this topic, that it is looking to identify non-LMOs that potentially come from two ends of the continuum that has LMOs in its centre. Robert posed the question “are there organisms so similar to those found in nature that they fall outside the Cartagena definition of LMO from the other end?”  Again reiterating my view that gene editing per se is not synthetic biology, I support Ilaria's suggestion [#9649] that the question of whether gene edited organisms fall within the LMO definition is not specific to synthetic biology. To me the focus of this exercise must be at the other end of the continuum that Robert identifies, "organisms developed using synthetic biology that are so different that they fall outside the definition of LMO under the Cartagena Protocol". In that regard, I agree with the many other participants who consider that there are no examples of synthetic biology organisms developed thus far that fall outside the LMO definition of the Cartagena Protocol. This includes the examples put forward by Carlos [#9640]; I agree with Robert [#9642] that the organisms Carlos describes are LMOs.

Best regards,
Louisa
posted on 2019-03-24 00:43 UTC by Dr. Louisa Matthew, Office of the Gene Technology Regulator, Australia
RE: Welcome message - synbio forum - topic 5 [#9650]
Dear all,

Thanks again to our moderator Casper for his guidance and the rest of Colleagues for their contributions and clever discussion

The Ad Hoc Technical Expert Group on synthetic biology in 2017 concluded that "most living organisms already developed or currently under research and development through techniques of synthetic biology, including organisms containing engineered gene drives, fell under the definition of LMOs as per the Cartagena Protocol." ( https://www.cbd.int/doc/c/aa10/9160/6c3fcedf265dbee686715016/synbio-ahteg-2017-01-03-en.pdf

As part of this AHTEG and still today I agree with this view, however most doesn’t mean all and the horizon scanning is very important in this regards, in fact there are interesting examples currently mentioned on this topic 5

On the other hand I would like to share and support the Jacks’ opinion in post [#9625]
“….. it might be found that such things pose little or no (unmanageable) risk, but defining them outside of the Protocol is to make an a priori decision that if no risk assessment was done that would be okay.”

I also agree with the approach [#9627] by Ricarda Steinbrecher and I would like specifically highlight her opinion that:
“If we agree on a wide scope and a broad interpretation then we will probably have all organisms currently produced through synthetic biology covered by the CPB. If not, then we need to clarify and ensure that they will be covered in other ways under the CBD to ensure environmental and human safety, including socio-economic considerations.”

I wish you all a nice weekend

Best regards,
Lazaro Regalado
posted on 2019-03-23 18:23 UTC by Mr. Lazaro Regalado, Cuba
RE: Welcome message - synbio forum - topic 5 [#9654]
Ossama [#9644] has said in a few words what is the essence of many interventions. Everything that is syn bio now and may be in the future is an LMO product of modern biotechnology. That is, unless for some reason it isn’t.

Echoing Sarah [#9648], Lazaro [#9650] and Ricarda [#9627] (and others), a potential solution is simply to define everything that might be synthetic biology as within the scope of the Protocol and thus subject to it. The alternative is what has lead to the European Court of Justice proceedings, and before that the High Court of New Zealand, arguing over the technicalities of words as strict lists. As I mentioned previously, with syn bio that future is to argue over the technicalities of what is living, organism, modification etc. For example, at what point does Nikolay’s example of virus-like macromolecular assemblies [#9605] transition from non-living machine to living organism?

One answer might be what Nikolay has already suggested, and that is the capacity to reproduce. Reproduction is a common characteristic of living things, but as the definition of one is contested. In any case, this boundary condition is dissatisfying to me. First, all nucleic acids find a way to reproduce given long enough or sufficiently large disseminations because of the diversity of DNA-dependent DNA polymerases, DNA-dependent RNA polymerases, RNA-dependent RNA polymerases and RNA-dependent DNA polymerases and the variety of DNA-DNA and RNA-RNA recombinases. Moreover, it relies on increasingly challenged views of viral infection range limitations (which are not as limited as symptom ranges) and overestimated confidence that the infection will be limited to the intended recipient organism. This is achievable in laboratories, but not in nature as we know from other examples of horizontal gene transfer.

If it is all but certain that reproduction at some level, either as the released entity or as an unintended but likely derivative made through recombination, will occur, then how is a protein-coated virus intended to deliver a nucleic acid cargo to the cells of a plant different from an aluminium-coated airplane intended to deliver a nucleic acid cargo formulated with a penetrant to fields of crop plants?

This I would argue is related to Ossama’s “parts and products of living organisms that result from Synbio” and why the emergence of open-air techniques, including gene drives and nucleic acids delivered through spray formulations (see e.g. Jim Thomas in Topic 6 [#9636]) is within our scope for discussion. It is my opinion that such things are best acknowledged to be within the operational scope of the Protocol (or at least CBD) rather than orphaned for future semantic debates.

All the best
Jack
posted on 2019-03-23 21:46 UTC by Mr. Jack Heinemann, University of Canterbury
RE: Welcome message - synbio forum - topic 5 [#9656]
Dear forum participants,

This topic has been discussed previously in online discussions and AHTEG meetings (2015 and 2017). Throughout this work the Global Industry Coalition (GIC) has maintained that living organisms already developed or currently under research and development through biotechnological tools are LMOs as defined by the Cartagena Protocol. Further, “synthetic biology” is within the scope of “biotechnology” as defined by the CBD, and “modern biotechnology” as defined by the Cartagena Protocol. This would include (currently conceptual) living organisms resulting from xenobiology, and exclude systems that are not living such as cell-free applications. Our more recent review for the 2019 GIC submission (https://bch.cbd.int/database/record.shtml?documentid=114285) did not change this view.

We emphasize that our position on this topic does not apply to applications of genome editing in plants; as we have stated previously, we do not believe that these belong in synthetic biology discussions. Genome editing can be used to achieve various outcomes in plants, with some outcomes comparable to existing biotech (LMO/GMO) plants, and others comparable to plants developed with conventional breeding tools. Where the product is an LMO, it is within the scope of the existing regulatory mechanisms that apply to LM crops developed using other biotechnological tools. Where the product is not an LMO, this is not a regulatory gap in the Cartagena Protocol or the international regulatory framework for LMOs/GMOs, rather, the same range of regulatory mechanisms should apply as for conventionally developed plants.
posted on 2019-03-24 00:51 UTC by Ms. Felicity Keiper, BASF
RE: Welcome message - synbio forum - topic 5 [#9658]
Dear colleagues,
First of all I would like to thank Maria and Caspar to facilitate jet another important discussion. By saying so I would like to reiterate the observation of Ilaria (#9649) that this online discussion and the submission by Parties are already providing useful contributions to the horizon scanning process of synthetic biology.

I would like to add to the discussion on certain applications of xenobiology that are synthetic biology, but I think that some applications are not LMOs. The applications I am talking about are Chemically Modified Organisms (CMO) (https://doi.org/10.1002/anie.201103010) that are modified with non-canonical nucleic acids or non-canonical amino acids (ncAAs). The latter are increasingly used in protein engineering (https://www.ncbi.nlm.nih.gov/pubmed/29781997). Both examples would fall under the operational definition of synthetic biology since they are “redesign living organisms”. But especially the CMOs that are modifies with non-canonical amino acids would not fall under the definition of LMO since they do not represent a novel combination of genetic material. Even thought this example might not have great impact on biodiversity (because they can only grow on specific media) I add it to the discussion for the sake of completeness.

Kind regards
Margret
posted on 2019-03-24 07:31 UTC by Dr. Margret Engelhard, Germany
RE: Welcome message - synbio forum - topic 5 [#9662]
I would like to thank Maria and Caspar for leading an interesting and important discussion.  As I am entering this discussion towards the end, most of the issues I was going to raise have already been covered, so I would like to raise a separate, though I believe connected, concern about this discussion. 

I am concerned that our discussions could introduce conflicting rules in relation to other United Nations treaties that also capture the tools, techniques and products of synthetic biology; particularly in relation to genome editing and whether or not these techniques and products fall under the definition of an LMO under the CBD.

The International Treaty on Plant Genetic Resources for Food and Agriculture states that “Contracting Parties shall develop and maintain appropriate policy and legal measures that promote the sustainable use of plant genetic resources for food and agriculture” which may include such measures as (* added for emphasis):

a) pursuing fair agricultural policies that promote, as appropriate, the development and maintenance of diverse farming systems that enhance the sustainable use of agricultural biological diversity and other natural resources;
*b) strengthening research which enhances and conserves biological diversity by maximizing intra- and inter-specific variation for the benefit of farmers, especially those who generate and use their own varieties and apply ecological principles in maintaining soil fertility and in combating diseases, weeds and pests;
c) promoting, as appropriate, plant breeding efforts which, with the participation of farmers, particularly in developing countries, strengthen the capacity to develop varieties particularly adapted to social, economic and ecological conditions, including in marginal areas;
*d) broadening the genetic base of crops and increasing the range of genetic diversity available to farmers;
e) promoting, as appropriate, the expanded use of local and locally adapted crops, varieties and underutilized species;
f) supporting, as appropriate, the wider use of diversity of varieties and species in on farm management, conservation and sustainable use of crops and creating strong links to plant breeding and agricultural development in order to reduce crop vulnerability and genetic erosion, and promote increased world food production compatible with sustainable development; and
*g) reviewing, and, as appropriate, adjusting breeding strategies and regulations concerning variety release and seed distribution.

Depending on how the CBD determines what constitutes a LMO and whether or not genome editing falls within this definition could very easily impact (b), (d), and (g) of the Plant Treaty.  I defer to the legal scholars in our group to reflect on which Treaty would take precedent if say the CBD determines that genome editing is an LMO and subsequent risk assessments are required, yet the Plant Treaty under (g) determines a different regulatory strategy for their release or believes that genome editing promotes the fulfillment of (b) and (d)? We have already seen a range of opinions in our discussions on this topic. Yet, there does not seem to be any coordination or cooperation with other UN Treaties on these topics.

I am not advocating one way or the other for a decision but rather calling for better coordination and cooperation with the Plant Treaty as they have begun to examine the impacts synthetic biology may have as it relates to the Plant Treaty (see attached report).

Kindest regards,

Todd Kuiken
posted on 2019-03-24 16:59 UTC by Mr. Todd Kuiken, North Carolina State University
RE: Welcome message - synbio forum - topic 5 [#9663]
Dear forum participants,

I agree with previous posts indicating that living organisms already developed or currently under research and development through modern biotechnology (including synthetic biology) are LMOs as defined by the Cartagena Protocol, as well as that systems which are not living (e.g. protocells) are excluded [e.g. #9619, #9621, #9637, #9649, #9656].

In terms of genome editing, and as already pointed out by various posts, it is clear that genome editing does not necessarily result in an LMO or synthetic biology [e.g. #9588; #9572; #9553; #9616; #9621; #9631], and that existing regulatory mechanisms for either LMOs/GMOs or conventionally developed organisms should be applied, depending on whether the organism is an LMO or not, respectively [#9656].

Kind regards, Delphine
posted on 2019-03-24 17:12 UTC by Ms. Delphine Beeckman, Belgian Biosafety Professionals
RE: Welcome message - synbio forum - topic 5 [#9665]
I believe when subsuming synthetic biology under the Cartagena definition of LMO we should be aware of the fact that the term and its definition is a legal one, i.e. one that not just mirrors scientific terminology.  I support Jack’s suggestion in # 9611 to distinguish between semantic and risk orientated understandings of LMOs as defined by the Cartagena Protocol (CP). I believe we should apply both (as lawyers do): semantics plus interpretation related to risk (which means: related to the purpose or “telos” of a term ). I understand Casper in # 9610, Ben David in # 9606, Birgit # 9639, Sarah 9648 and others to have this in mind too. This means that the definition of LMO although at first sight narrowly phrased may be expanded to cover processes/products of SynBio that were originally not meant to be covered (most often because they were not known before). Or that we suggest that some products are clearly not covered by the definition which would then necessitate deliberations of the AHTEG and finally the Cartagena contracting parties on whether the scope of the CP should be enlarged. Vice versa the CP LMO definition might also be narrowed down to exclude certain processes/products from oversight because they are considered to pose not any or only negligible risk.
On this basis some of the examples appearing in this discussion could be tested for being or not covered by the CP LMO definition , having in mind that its core elements are “living”, “modified” and “organism”, “novel combination of genetic material” and “application of […] In vitro nucleic acid techniques, including recombinant deoxyribonucleic acid (DNA)”. [note: I am not a scientist so apologies if I got the examples wrong]:
1. Modification of an organism using only protein reagents (Mega nucleases, ZFN, TALEN) introduced into the cell without involving the modification of the genotype (cf Nicolay # 9605): can this be included in the term “in vitro nucleic acid techniques”? Nicolay says: no. Helmut (# 9623) may be too. If that is correct: I would expect that scientists discuss possible risks and discuss if the scope of CP should expanded.
2. Chemically modified organism that do not carry novel combination of genetic material (cf Margaret # 9658): clearly not included. Same issue concerning risk and expansion of scope.
3. Virus-like macromolecular assemblies (cf Nicolay # 9605): Modified viruses seem to be covered by the LMO definition (right?), so why not also virus like assemblies? Only if they can (or might) replicate inside cells?
4. Bioparts, not replicating protocells (see Ossama # 9644): clearly not covered, but once more the question of risk and expansion of scope emerges.
5. Genome editing: if understood to be cisgenesis i.e. not involving transgenes (am I right?): following the recent judgment of the EU Court of Justice gene edited organisms if triggered through non-traditional mutagenesis would be included in the term. “Novel combination of genetic material” is also one that does not involve transgenes. Luz (# 9608) seems to see this differently, maybe also Felicity #9656.
6. Introduction of XNA into an organism (cf Boet # 9619): it’s clearly a novel combination of genetic material, but it is not recombinant DNA. So, does the “application of in vitro nucleic acid techniques” reach beyond the modification of DNA? If so, XNA is covered. This result would be “teleologically” supported if one could show that an XNA  modified organism poses risks.
7. Minimal cells (eg Bob # 9642): should they be excluded from scope because they are not “really “ living but just a chassis for further use which will then be a LMO?
8. An organism that was treated by application of in vitro nucleic acid techniques but is entirely identical to its naturally occurring genome (cf Nikolay # 9624): Is it a LMO? Nikolay and others (Bob # 9628) seem to say no. But there was application of in vitro nucleic acid techniques, and the organism  was modified in a procedural sense, even though not in effect. I would look at possible risks here and decide the question accordingly.
9. Entirely newly synthesized organisms: this has not been achieved so far but may happen in the near future: Would such organism be a LMO in the CP-sense? LMO means modification of an organism. A completely new one is this not, or is it?

Todd (# 9662) raised one very important additional issue, i.e. the systematic context of the CP definition of LMO. The CP has risk assessment and management in mind while the ITPGR is orientated to encourage use and see to benefit sharing. He is right to be concerned about possible conflicts such as if the scope of the CP is broad it may hinder the bringing on the market of modified plants. It should therefore be ensured that the term LMO is defined having potential risks in mind. If that is the case this would be compatible with the ITPGR. The same problem arises with regard to the Nagoya Protocol (NP) which organizes benefit sharing beyond the scope of the ITPGR, and should be solved accordingly. It should be added that the trigger term for the application of both ITPGR and NP is not LMO but genetic resource. The genetic modification of organisms is only one case of the utilization of genetic resources and thus of the right of provider states to claim benefit sharing.
Incidentally: the objective of the CBD is among others sustainable use of genetic resources: Has this ever been seriously discussed outside the ITPGR as a concern? In relation to the CP the question could be posed if any objective of modifying organisms is legitimate if it is only ensured that side-effects on human health and the environment are prevented. For instance, should transgenes accelerating growth and size of cattle and pork be indefinitely be transferable to them making animals pure machines of meat production without any concern for their dignity? If these machines conquer the world agriculture is it sustainable if all other wild relatives are pushed off and die out?
Sorry for these digressions but I share Todd’s concern that we should be aware of broader contexts.
Best wishes
Gerd
posted on 2019-03-24 18:05 UTC by Mr. Gerd Winter, University of Bremen
RE: Welcome message - synbio forum - topic 5 [#9667]
Dear Maria, Casper, dear colleagues,

My participation in our current forum comes towards the end of topics 5 & 6 and I will introduce myself. I am Maria Mercedes Roca, a plant pathologist/biotechnologist, formerly a regulator for the Government of Honduras and a member of the Risk Assessment and Risk Management AHTEG, until I moved to Mexico, where I also participate actively with biotech/biosafety issues through CIBIOGEM.

I thank our moderators and all participants for a very important, lively and yes, still difficult discussion on how we should define synthetic biology and thus,  specifically on  topic 5, following Casper’s instruction to  “Consider whether any living organism developed thus far through new developments in synthetic biology fall outside the definition of living modified organisms as per the Cartagena Protocol”.
Simply put, yes, all living organisms developed so far through synthetic biology are LMOs. We have discussed this topic on previous for and seem to be going round in circles with the arguments.

We all agree that to move on,  it is useful to have an operational definition of synthetic biology. Yet,  as Jim  (#9638) and many others point out,  the definition of synthetic biology has as it stands today,  a lot of “fuzziness around the edges” that creates confusion today, and  may well lead to important errors for future oversight. This is especially true for policy development in developing countries such as Central American and Andean countries, with little resources for biosafety policy development and implementation.

With the exception of possibly Colombia, policy arguments regarding biotechnology, still go round issues of GMOs/transgenics and not synthetic biology, a largely unknown and misunderstood term, even among  local scientists and regulators. 
I applaud Jack’s (#9625)  and Ricarda’s  (# 9627) attempts to “think outside the box” when considering if products of synthetic biology are (or not) LMO’s,  and especially like Bob’s  (#9628) comment – also thinking outside the box” of a “continuum” regarding the definition of an LMO.
I strongly support Luz’s statement from Colombia (# 9608)  that the products  and applications,  and not the tools of genetic engineering, should guide regulatory oversight. I also agree with Felicity (# 9572), Piet (#9588), Boet (#9619), Jim (#9638), Delphine (#9632), Christoph (#9635), Rodrigo (# 9637) and many others,  that products of synthetic biology so far developed are LMOs and thus should come under the Cartagena Protocol . Furthermore, risk assessment for LMOs so far developed, including products of genome editing is adequate and does not require further guidance at this point.  

Finally, I strongly agree with Swantje (#9631), Jenna (#9632), and others, that genome editing is not the same as synthetic biology, as both terms are too broad.
I look forward to discussions on Topic 7.

Best regards,

Maria Mercedes
posted on 2019-03-24 20:05 UTC by Dr. Maria Mercedes Roca, CIBIOGEM, Mexico
RE: Welcome message - synbio forum - topic 5 [#9666]
Dear Colleagues.

First I would like to thank Maria and Casper for moderating the discussions of these topics concerning synthethic biology.
The discussions are really helpfull and have allready given allot of insight around the topic and given allot of usefull knowlegde.

Following topic 5, I agree that most organism derived through Synthethic Biology at current stage, fall under the scope of the defintions of LMOs according to the Cartegena protocol. I can understand how some can find it difficult to create the right RA for organsims containing synthetic parts, but in my opinion, it is most important to still do a case by case RA, and be aware of the extra risk some organsim with syntethic elements can have. Eg that some organism may behave different i nature compared to a LMO where you almost can predict the survival and effect of natural habitats.

However we have experienced one example where a organism was derived completely from synthethic DNA, where a core of virus proteins were used to encapsule the DNA, and used to fuse the DNA with bacteria. To this extenct of organism i find it difficult to find the connection of these kinds of "organism" as being under the scope of the Cartegena protocol.

Again thank you for these usefull disucissions, its a pleasure to follow.

Kind regards
Selina Hansen, Danish Enviromental Protection Agency.
posted on 2019-03-24 19:27 UTC by Ms. Selina Kruuse Hansen, Danish Enviromental Protection Agency
RE: Welcome message - synbio forum - topic 5 [#9669]
Dear colleagues,

I echo the belief from several  (#9619, #9621, #9629, #9631, #9633, #9635, #9649, #9650, #9656, #9663, etc.) that living organisms from current and foreseeable applications of Synthetic Biology fall within the definition of LMOs as per the Cartagena Protocol.

I'd also like to repeat that genome editing tools do not necessarily result in synthetic biology. There are different possible outcomes which may be an LMO or similar to conventionally bred improvement. I agree that risk assessment and management is on a case-by-case basis. 

Looking forward to further discussions.

Best regards, Lúcia
posted on 2019-03-24 21:02 UTC by Ph.D. Lúcia de Souza, PRRI - Public Research and Regulation Initiative/ANBio (Associação Nacional de Biossegurança - Brazilian Biosafety Association)
RE: Welcome message - synbio forum - topic 5 [#9670]
Dear Casper,
Thanks for your guiding of this forum, and for your steadfastness in bringing the discussion back to the question on the table, i.e.: are there living organism developed through synthetic biology that fall outside the definition of LMO?
As many colleagues have said, this all comes back to the definition of GMO.
Before sharing some observations in this perspective, first a quick correction: in your presentation of the definition LMO you placed the phrase “that overcome natural physiological reproductive or recombination barriers and that are not techniques used in traditional breeding and selection” after the sentence “Fusion of cells beyond the taxonomic family”. That is incorrect. The phrase “that overcome …………” refers to both parts a. and b. of the LMO definition. The lay out of the definition reflects that.
Some of the observations in this forum about DNA and RNA were also discussed during the negotiations, as a result of which the definition refers in general to nucleic acid, covering DNA and RNA. The negotiations on the definition aimed to provide clarity for existing and future developments. In this respect I warmly support Jim Louter’s [#9638] tribute to the scientists and negotiators who crafted the LMO definition..
Bearing in mind that the operative term in Casper’s question topic is “developed” (i.e. organisms that have actually been produced), I concur with Ilaria Ciabatti [#9649], Boet Glandorf (#9619), Motlalepula [#9615], Bob Friedman, JCVI [#9628], Swantje Strassheim, [#9631]and others that organisms developed through synthetic biology, currently and in the foreseeable future, fall under the LMO definition, and that therefore the case by case risk assessment will apply.
The posts of Helmut Gaugitsch [#9623], Nikolay Tzvetkov [#9624], Selina Hansen [#9666], Jack Heinemann [#9609], Guy Reeves, [#9622] and others illustrate that it will indeed be good to keep an eye of developments in this field.
If at one point it is established that certain organisms developed through SynBio do not fall under the LMO definition, then comes the question whether there are scientific reasons related to the objectives of the CBD that would warrant to bring those organisms under regulatory oversight (see also the post of Birgit Winkel, [#9639]).
While still a bit sour that the German team defeated the Dutch team tonight, I wish you all a great end of the weekend

Piet
posted on 2019-03-24 22:20 UTC by Mr. Piet van der Meer, Ghent University, Free University of Brussels, Belgium
RE: Welcome message - synbio forum - topic 5 [#9672]
Dear All,

In accordance with the precautionary approach contained in Principle 15 of the Rio Declaration on Environment and Development, the objective of this Cartagena Protocol on Biosafety  is to contribute to ensuring an adequate level of protection in the field of the safe transfer, handling and use of living modified organisms resulting from modern biotechnology that may have adverse effects on the conservation and sustainable use of biological diversity, taking also into account risks to human health, and specifically focusing on transboundary movements. http://bch.cbd.int/protocol/text/

"Living modified organism" means any living organism that possesses a novel combination of genetic material obtained through the use of modern biotechnology;
http://bch.cbd.int/protocol/text/

"Living organism" means any biological entity capable of transferring or replicating genetic material, including sterile organisms, viruses and viroids;
http://bch.cbd.int/protocol/text/

"Modern biotechnology" means the application of:
a.In vitro nucleic acid techniques, including recombinant deoxyribonucleic acid (DNA) and direct injection of nucleic acid into cells or organelles, or
b. Fusion of cells beyond the taxonomic family, that overcome natural physiological reproductive or recombination barriers and that are not techniques used in traditional breeding and selection;
http://bch.cbd.int/protocol/text/

Ad Hoc Technical Expert Group on synthetic biology in 2017 concluded that "most living organisms already developed or currently under research and development through techniques of synthetic biology, including organisms containing engineered gene drives, fell under the definition of LMOs as per the Cartagena Protocol."
( https://www.cbd.int/doc/c/aa10/9160/6c3fcedf265dbee686715016/synbio-ahteg-2017-01-03-en.pdf

In this regard, I suggest to consider the objective of Cartagena Protocol to contribute to ensuring an adequate level of protection use of living modified organisms resulting from modern biotechnology that may have adverse effects on the conservation and sustainable use of biological diversity. Hence, we need an operational definition of synthetic biology to help to clarify ad Hoc Technical Expert Group on synthetic biology.

I understand living modified organism possesses a novel combination obtained through the use of modern biotechnology. Taking account this, protocells (https://pubs.acs.org/doi/abs/10.1021/ja808018y), genome editing (https://www.nature.com/articles/nbt.2501  https://www.sciencedirect.com/science/article/pii/S0734975014001931), artificial cells  (https://www.sciencemag.org/news/2018/11/biologists-create-most-lifelike-artificial-cells-yet   https://royalsocietypublishing.org/doi/full/10.1098/rsfs.2018.0046 ) fall outside the definition of living modified organisms as per the Cartagena Protocol.

I agree with the following experts #9663; #9619; #9621; #9637; #9649; #9656; #9588; #9572; #9553; #9616; #9621; #9631; #9656; #9631; #9632 and #9667.

Thanks so much,

Dr. Marina Rosales Benites de Franco
posted on 2019-03-24 22:47 UTC by Ms. Marina Rosales Benites de Franco, National University Federico Villarreal
RE: Welcome message - synbio forum - topic 5 [#9673]
Dear forum participants,

Thank you for your interventions. This topic is now closed for comments.

Secretariat
posted on 2019-03-25 01:00 UTC by Mr Austein McLoughlin, Secretariat of the Convention on Biological Diversity