Past Activities 2019-2020

Welcome to the Open-Ended Forum on Synthetic Biology. In its decision 14/19, the Conference of the Parties decided to “extend the Online Forum on synthetic biology taking into account the work on risk assessment under the Cartagena protocol, to support the deliberations of the AHTEG".
To give effect to the COP’s decision, 7 topics have been selected for discussion under this forum.
The topics are:
Topic 1: New technological developments in synthetic biology since the last meeting of the Ad Hoc Technical Expert Group
Topic 2: Recommend options for carrying out the regular horizon scanning, monitoring and assessing of developments referred to in para 3 of decision 14/19
Topic 3: Review of the current state of knowledge
Topic 4: Possible impacts of synthetic biology applications that are in early stages of research and development on the three objectives of the Convention
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
Topic 6: Sharing of experiences on detection, identification and monitoring of organisms, components and products of synthetic biology
Topic 7: Relationship between synthetic biology and the criteria set out in decision IX/29
This forum is co-moderated by Mr. Casper Linnestad from Norway (Norwegian Ministry of Climate and Environment), and Ms. Maria de Lourdes Torres from Ecuador (Universidad San Francisco de Quito).
During the first week, topics 1 and 2 will be open for discussion. The second week will consider topics 3 and 4, the third week topics 5 and 6 and the fourth week will be for discussion on topic 7.
Each moderator will take the lead in supporting discussions under one of the two topics each week, and they will work together for the moderation of topic 7. For the topics that will be open for discussion this week, Ms. Torres will moderate topic 1, and Mr. Linnestad topic 2.
We now request participants to the Online Forum to provide submissions on topics 1 and 2 for the next week. Discussions under these topics will close at 1 am GMT on Sunday 10th March. You can see the schedule for discussions on other topics on the forum page.
We suggest that submissions are always as specific as possible to facilitate the analysis of the information as well as the interpretation of each post from other forum participants. We kindly request you to refrain from commenting on topics that are not yet open for discussion, and only provide inputs to the discussions that are open on each particular week.
In addition, if you would like to respond to another post, we would like to encourage you to mention the # of post you are responding to, for example: “in response to post # XXXX, …”. This will also facilitate the understanding of others.
Please post your responses under this thread. We also advise you to review the general guidelines for the forum on the main webpage of the discussion.
We thank you for your participation in this forum, and we look forward to an interesting discussion.
Sincere regards
Casper Linnestad and Maria de Lourdes Torres
Co-Moderators

Dear all

On behalf of my country, Gabon, I want to congratulate both all our moderators for this online forum on synthetic biology. It will be my pleasure to participate once again to discussions.

Best regards

Dr. Jean Bruno MIKISSA

Dear all,
Thanks to the Secretariat for the open-ended forum and congratulations to both moderators. I hope this forum will be as clear and useful the issue of SynBio requires. It will be a pleasure to participate.
Sincerely yours,
Pedro Rocha

Thanks to the Secretariat for this new open-ended forum and congratulations to Casper an Maria de Lourdes. It will be a pleasure to participate once again in this forum.

Best regardas,

Maria Andrea

Dear All,

I am pleased to participate in the interesting forum on synthetic biology. This will be first interaction from my country on the issue. Looking forward.
My appreciation to the secretariat and moderators.
  Nada Babiker Hamza
  Sudan

Looking forward to productive discussions. We are most happy to engage.

Dear all,

Thank you to the moderators and to the Secretariat. We at the University of California, Berkeley look forward to engaging with you all and contributing to productive discussions via this forum.

Regards,

Valeri Vasquez

Dear colleagues,
My name is Julio Valdivia, Chair of the Bioengineering and Chemical Engineering Department at Universidad de Ingenieria y Tecnologia UTEC, Lima Peru. Thanks for being part of this discussion. My country is one of the most biodiverse areas in the world and obviously, we are concern about the possible damage that could be generated by synthetic biology, however, it is not the real issue, the absence of fluent and rational regulations is the real one. We have to generate new strategies to keep safe diversity together with the advance in biotech. It is a huge wave, depend on how we want to receive it.

Dear participants,
I’m Lázaro Regalado from Cuba, Biodiversity and Biosafety Department / Office of Regulation and Environmental Safety.
I would like to congratulate Moderators Maria and Carper and the Secretariat  as well as the rest of moderators in advance for this new round of online deliberations that seek to find answers to these 7 topics in order to support the deliberations of the AHTEG.
I hope to participate to fulfill our mandate and collaborate with this remarkable group of experts.

Respectfully,
L. Regalado

Dear Colleagues,

This is Wei Wei from China. I work in the Institute of Botany, Chinese Academy of Sciences and currently am interested in risk assessment of synthetic biology using lessons and experiences learned from  genetically modified organisms.

Regarding of the discussion topic 1, I do not see any new technological progress since 2017 (Please correct me if I am wrong). There are definitely plenty of achievements in the application of established techniques in new organisms. For example, the application of CRISPR-Cas9.

However, there is ongoing discussions on the use of synthetic biology in biodiversity conservation and their interaction. For example, to extend the gene diversity for better adaptation to changing climate, to eliminate pests and deceases of endangered species. It is very interesting and closely relevant to the convention. I think that it is also good to discuss and comment on this trend of progress and its impacts.

I look forward to a fruitful discussion.

Sincerely

Wei

Dear Colleagues,
I am pleased to participate in this interesting forum on synthetic biology. Many thanks to the Secretariate and Co-chairs for moderating.
Looking at the 2017 AHTEG report I agree with Wei Wei that I don`t see new technological approaches when comparing p. 15 of those report. At the same time there many achievements in established techniques. Some of them are more fundamental, but at the same time interest to such technicues and continuity of such studies, development of more complex constructs, a lot of workshops and trainings that were held and ongoing on this topics (e.g. http://mammalian-synbio.org/2018,
https://www.aiche.org/sbe/conferences/metabolic-engineering-conference/2018 https://www.pioneeringminds.com/phil-stuart-2-2/ and many others ongoing in 2019)., as well as Start-up proposals, suggests that soon we can expect practical applications and need to screen this new more complex developments.
There are also discussions on projects on rodents elimination.
First successful gene drive in mammals https://www.the-scientist.com/news-opinion/first-successful-gene-drive-in-mammals-65367.

Looking forward for contribution from colleagues.
Best wishes,
Galina

Dear Colleagues,
My name is Nikolay Tzvetkov and I work at the Bulgarian Ministry of Environment and Water. I also had the honour to participate at the AHTEG meeting in 2015 and in 2017.

First I would like to congratulate the moderators and wish them success in their work.

Regarding the new technological developments in the field of Synthetic biology since the last AHTEG meeting I think the replies to Notification 2018-103 provide good overview of some of the most important technological developments since December 2017 (https://bch.cbd.int/synbio/submissions/).

Regarding the specific questions posed by the moderator I would like to draw your attention to the following papers:
• Which concrete applications of genome editing related to synthetic biology have been made recently?
1. In vivo CRISPR editing with no detectable genome-wide off-target mutations, Akcakaya et al., Nature 561 (7723), 416-419
A highly sensitive strategy that can robustly identify the genome-wide off-target effects of CRISPR–Cas nucleases in vivo is described. The paper shows show that appropriately designed guide RNAs can direct efficient in vivo editing with no detectable off-target mutations.
2. Predictable and precise template-free CRISPR editing of pathogenic variants, Shen et al., Nature 563 (7733), 646-651
The paper presents an approach for precise, template-free genome editing.
3. Domestication of wild tomato is accelerated by genome editing, Li et al., Nature Biotechnology 36, 1160-1163
4. De novo domestication of wild tomato using genome editing, Zsogon et al., Nature Biotechnology 36, 1211-1216
5. Rapid improvement of domestication traits in an orphan crop by genome editing, Lemmon et al., Nature Plants 4, 766-770
Those three papers demonstrate that gene editing can be used for fast and efficient domestication of wild plants. This is relevant not only for the first two objectives of the Convention but also for the third one.
6. Predictable and precise template-free CRISPR editing of pathogenic variants, Shen et al., Nature 563 (7733), 646-651
The paper presents an approach for precise, template-free genome editing.

• What new developments are known—related to the synthesis of whole genomes and chromosomes—that could have a real impact on biodiversity?
1. Creating a functional single-chromosome yeast, Shao et al., Nature 560 (7718), 331-335
2. Karyotype engineering by chromosome fusion leads to reproductive isolation in yeast, Luo et al., Nature 560 (7718), 392-396
Those two papers present creation of yeast with altered chromosome number. This demonstrates the feasibility of chromosomal engineering. Such organisms if released into the environment will be sexually incompatible with their wild relatives and this be safer in some respects.
3. Biotechnological mass production of DNA origami, Praetorius et al., Nature 552 (7683) 84-87
This paper presents a method for scalable productions of large DNA molecules of arbitrary sequence that can be used for production of complex nanostructures.
4. Evolution of a designed protein assembly encapsulating its own RNA genome, Butterfield et al., Nature 552 (7685), 415-420
The paper presents the development of synthetic nucleocapsids that can package their own RNA genome. This is an example of de novo design of virus-like structures.
5. Synthetic glycolate metabolism pathways stimulate crop growth and productivity in the field, South et al., Science 363 (6422), eaat9077
The study demonstrates that engineered metabolic pathways can affect and improve the agricultural properties of plants. Such metabolic engineering approaches will be greatly facilitated by the ability to synthesize and introduce into the cell large fragments of DNA or whole chromosomes.

• Can we report on the progress of engineered gene drives in sexually reproducing organisms? Do we know about new applications of this technology?
1. Super-Mendelian inheritance mediated by CRISPR–Cas9 in the female mouse germline, Grunwald et al., Nature 566 (7742), 105-109
A gene drive system in mammalians is described.
2. A CRISPR-Ca9 gene drive targeting doublesex causes complete population suppression in caged Anopheles gambiae mosquitoes, Kyrou et al., Nature Biotechnology 36, 1062-1066
The paper describes an efficient gene drive that spreads rapidly in a population of caged mosquitoes to the point of total population collapse.
3. Small-Molecule Agonists of Ae. aegypti Neuropeptide Y Receptor Block Mosquito Biting, Duvall et al., Cell 176 (4), 687-701
Although not utilizing synthetic biology, the paper demonstrates how small-molecule compounds can be used to control disease vectors through altering their behaviour.

Best Regards,
Nikolay

Dear all, I agree with Wei Wei and Galina regarding new technologies vs achievements. I think it boils down to at what point an edited or modified organism become a product of synthetic biology. 

Marie

Dear all,
First of all I would like to send my congratulations to the two moderators.
Second, I would like to answer to Wei Wei (post 9363) and Galina (Post 9364). I also think that not only the new development is of importance, but also the way it will be used and introduced into the environment needs to be monitored.
The difference of synthetic biology and genetic modification is not the technic that is used or the amount or the depth of changes in the genome, but the idea behind. Synthetic biology aims to use life as a modular construction system without taking into account that organisms play a certain role in the ecosystem.
Regards
Birgit

--THIS MESSAGE IS POSTED ON BEHALF OF THIS TOPIC'S MODERATOR MS. MARIA DE LOURDES TORRES--

Dear Forum Participants,

Thank you for your welcoming messages to this forum, Topic 1: New technological developments in synthetic biology since the last meeting of the Ad Hoc Technical Expert Group.

I would also like to thank to all participants who until now have shared substantive information that enriches this forum.

In order to be able to follow all the interventions in a proper way, please post comments replying to the original thread with the opening message.

If we create separate threads, it will be hard to follow our conversation. Thanks in advanced.

I hope that in the following days we might continue to share valuable information regarding the topic of this forum

Kind Regards,

Maria

Hello,

I am Louisa Matthew from the Office of the Gene Technology Regulator, the Australian agency responsible for assessing applications to work with LMOs in contained facilities and the open environment.

I echo the points made by Wei Wei [#9363], Galina [#9364], Marie [9371] and others. I consider that, while different applications may have progressed, the underlying technologies have not changed since the 2017 AHTEG report. An Australian example is the development of canola genetically modified for omega-3 oil content, which has been described by some as synthetic biology. This involved insertion of a multi-gene pathway of seven genes sourced from yeast and several microalga species, using standard gene technology methods, to create a new oil profile. In February 2018 Australia’s Gene Technology Regulator approved commercial cultivation of this canola, and the full risk assessment to support this decision is available here: http://www.ogtr.gov.au/internet/ogtr/publishing.nsf/Content/DIR155

Kind regards,
Louisa

My name is Margret Engelhard and I work for the Federal Agency for Nature Conservation in Germany and was an active member of the AHTEG in 2015 and in 2017.

In addition to the very helpful substantial previous posts, new developments in synthetic biology that could be relevant to the objectives of the convention also include:

1.) GM-Virus applications in agriculture including those that are transmitted by insects (HEGAAs)
2.) Re-domestication of crop plants
3.) Xenobiological developments
4.) Development of GM – mammals are currently experiencing a huge boost, including first gene-drive in mammals
5.) GM applications in the nature conservation sector

In the following, I would like to further elaborate on these points:

Add 1.) Environmental release of GM-Virus is already ongoing. Permit for environmental release of GM-Viruses in Citrus tree to confer resistance to the bacterial disease known as citrus greening has been applied for in the US (https://www.aphis.usda.gov/brs/aphisdocs/17_044101r_CTV_dEIS.pdf). Viruses have potentially elevated propagation characteristics compared to e.g. GM-plants and thus need adequate scrutiny.

On special concept in this area called HEGAA seeks to utilize GM-Viruses that are transmitted by insects to crop plants in order to perform genetic engineering in the field (Reeves et al. 2018 DOI: 10.1126/science.aat7664). HEGAAs take a laboratory in the field approach to a new level. HEGAAs potentials for spread might be similar or even outpace those of gene drives (Simon et al. 2018 DOI: 10.1126/science.aav7568).

Add 2.) The use of genome editing for re-domestication as seen for tomato (Zsögön et al 2018; doi.1038/nbt.4272; also mentioned in post #9366). This approach represents a novel chality of intervention as domestication is performed with only a few steps in an extremely short time frame. Potentially known and unknown advantages but also adverse traits might be retained in the newly domesticated variety, with potential side effects to biodiversity and human health.

Add 3.) Xenobiological synthetic biology applications are further advancing and aim to expand the framework of natural chemistries within living cells through the incorporation of non-natural building blocks (Diwo and Budisa, 2018 DOI: 10.3390/genes10010017; also mentioned in post #9373). Here specific questions for example regarding the degradability of non-natural building blocks in the environment need to be tackled.

Add 5.) As also mentioned in post #9363, some research threads of synthetic biology focus on nature conservation applications. Besides challenges to the risk assessment of these applications due to our limited experience of the genetic alteration of wildlife, more general questions with respect to nature conservation need to be considered. What are the long term effects of a (gen-)technology-oriented approach to nature conservation that focuses on symptomatic problem solving instead of remedying the causes? Does the use of genetic engineering not change current nature conservation categories? For example, is a genetically modified (originally protected) organism still worth protecting? The further the synthetic biology project progresses to create artificial nature, the greater will be the influence on current concepts of nature (and nature protection). The discussion on the admissibility of genetic engineering in nature conservation, must therefore focus on the search of the border between the nature and biodiversity that has naturally emerged and a (genetically edited) nature that has been (man) made. The question whether the use of genetic engineering in nature conservation applications is a critical transgression of this border, needs to be addressed at the CBD level, also making use of the knowledge and taking into account the specific circumstances of IPLCs.

Dear colleagues,

I am Xu Jing from Chinese Research Academy of Environmental Sciences (CRAES). My pleasure to participate into this online discussion. I would like to echo Prof. Wei Wei's intervention, that there is no progress being made since 2017, although the publications and articles regarding to Synbio have been published. I think this would be a fact that we have to notice while we start the discussion. Thank you.

Dear colleagues,
My name is Maryna Bahdanava and I represent the National Co-ordination Biosafety Centre (NCBC) of Belarus.
I want to express sincere thanks for the possibility to participate in this Forum. I would like to send my congratulations to the two moderators.
I fully agree with Mr. Jeremy Sweet [# 9378] and Mr. Andrew Roberts [# 9372] on the need to clarify the concept of "synthetic biology". I think it is very important to specifically write where biotechnology ends and synthetic biology begins. Otherwise, most modern developments can be attributed to synthetic biology.
With regard to specific issues, I think special attention should be paid to the following articles:
S. Hoshika et al., “Hachimoji DNA and RNA: A genetic system with eight building blocks,” Science, 363:884–87, 2019.
Grunwald HA, Gantz VM, Poplawski G, Xu XS, Bier E, Cooper KL. Super-Mendelian inheritance mediated by CRISPR-Cas9 in the female mouse germline. Nature. 2019 Feb; 566 (7742): 105-109. doi: 10.1038 / s41586-019-0875-2. Epub 2019 Jan 23.

Dear colleagues,

Greetings! My name is Claudia Vickers. I work at CSIRO, Australia's Federal research agency, and also at The University of Queensland, Australia. I am pleased to contribute to this Forum and I look forward to to coming discussions.

With regards to advances in synthetic biology since the previous Forum, I agree with numerous other posts that there have not necessarily been material advances in synthetic biology technologies since the 2017 AHTEG report. For example, we have been able for some time to synthesize whole chromosomes, introduce them into cells, edit on the genome, build metabolic pathways, control regulatory circuitry, etc. etc.

I also agree that applications of synthetic biology have progressed and there are many new examples. This is quite normal for new technologies as their uptake increases and creative human beings start applying these new technologies to their own questions and problems.

This variation in opinion is probably based in the observation of Maryna (#9382) that there is not necessarily universal agreement on the definition of synthetic biology. However, there is reasonably high similarity between most commonly-used definitions nowadays.

In Australia we have released in September last year the Australian Council of Learned Academies' report on synthetic biology, entitled 'Synthetic Biology in Australia: an outlook to 2030'. This report was commissioned by the Office of the Chief Scientist of Australia and is part of a horizon scanning series that helps inform policy development. I was on the Expert Working Group that prepared the report, and after very extensive deliberations (which are summarised in the report), we settled on the following definition: ‘the rational design and construction of nucleic acid sequences or proteins – and novel combinations thereof, using standardised genetic parts’. This is a fairly broad definition and very process-based, as are almost all definitions, which is appropriate for a field that is applied by nature. You can find the report here: https://acola.org.au/wp/sbio/

On the basis of definitions such as this, which as noted are becoming fairly standard, it is fair to say that technical aspects of synthetic biology have not changed dramatically since the previous Forum. So perhaps, we should focus the topic on new applications of synthetic biology in order to stimulate useful discussions?

Best regards,
Claudia

Dear all,

My name is Boet Glandorf and I work at the Dept. of Gene Technology and Biological Safety, which is part of the Dutch National Institute of Public Health and the Environment. I also had the honour to be a member of the AHTEG in 2015 and in 2017.

First of all, my congratulations and best wishes for our moderators.

Regarding the topic at hand, new technological developments in the field of synthetic biology since 2017, I too would like to refer to the replies to Notification 2018-103 (https://bch.cbd.int/synbio/submissions/).

Developments in research on synthetic biology have been documented in literature, newsletters, websites (for example from start-up companies) and through iGEM competitions. These developments overlap with those in genetic modification. Therefore, I would rather refer to developments in modern biotechnology, instead of to developments in synthetic biology.

Some reports since 2017 highlight new developments and concrete applications of modern biotechnology, expected timelines and the consequence for risk assessment. These are:
-Preparing for future products of biotechnology (2017). Report of the National Academy of Science
https://www.nap.edu/catalog/24605/preparing-for-future-products-of-biotechnology
-Assessment of human health and environmental risks of new developments in modern biotechnology - a policy report (2018). RIVM report (Hogervorst et al.).
https://www.rivm.nl/publicaties/assessment-of-human-health-and-environmental-risks-of-new-developments-in-modern-biotechnology
The RIVM report is based on three reports commissioned by RIVM that describe new developments in red, white and green biotechnology, among others. References can be found in the report. 

A Dutch newsletter on new developments and applications in modern biotechnology and synthetic biology (titled ‘the SynBiOnt’) is published by our department on a regular basis and is now integrated our biotechnology site  https://biotechnologie.rivm.nl/
Please find below a summary of the main publications in the newsletter and on our website since 2017, which give an impression on the progress made in modern biotechnology.

Development of new techniques, new applications
- GM bacteria kill tumors
https://www.ncbi.nlm.nih.gov/pubmed/28179508
- Genetic firewall
http://biorxiv.org/content/biorxiv/early/2017/03/09/115493.full.pdf
- Reorganizing metabolic pathways in yeast
http://www.pnas.org/content/113/52/15060.full.pdf
- Biosensors
http://www.nature.com/nbt/journal/v35/n4/pdf/nbt.3791.pdf
- Synthetic spider silk
https://www.technologyreview.com/s/541361/spinning-synthetic-spider-silk/
https://www.technologyreview.com/s/603817/synthetic-spider-silk-for-sale-in-a-314-necktie/
- DNA origami
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5396133/
- Gibson assembly technique
https://www.nature.com/nbt/journal/vaop/ncurrent/pdf/nbt.3859.pdf
- Bacterial blight resistance in rice
http://dx.doi.org/10.1038/nature22497, http://dx.doi.org/10.1038/nature22371
- Artificial Intelligence
http://www.sciencemag.org/news/2017/07/new-breed-scientist-brains-silicon
- Nitrogen fixing bacteria
https://www.forbes.com/sites/susanadams/2017/09/14/new-venture-aims-to-make-crops-produce-their-own-nitrogen-fertilizer/#3c2f79951db0
http://www.press.bayer.com/baynews/baynews.nsf/id/Bayer-Ginkgo-Bioworks-forces-sustainable-agriculture-forming-company-USD-million-Series-A
- Skin transplantation
https://www.nature.com/articles/nature24487.epdf
- New chemical compound produced by E. coli
https://www.nature.com/articles/nature24996
- synthetic cell kills tumor
http://onlinelibrary.wiley.com/doi/10.1002/adhm.201701163/epdf
- Reconstruction horse pox virus and dual use
http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0188453
- Biosensors in medicine
https://globalgenes.org/raredaily/synlogic‐reports‐positive‐results‐in‐early‐stagestudy‐
of‐treatment‐for‐ucd/
http://blogs.plos.org/synbio/2018/01/29/engineered‐bacteria‐to‐detect‐gutinflammation
- Development of synthetic cell
http://www.nature.com/articles/s41467‐018‐03926‐1.pdf https://www.cogem.net/index.cfm/nl/publicaties/publicatie/het‐bionano‐avontuur‐bouwen‐aan‐delevende‐cel
-       CRISPR and broadly applicable blood cells
http://embomolmed.embopress.org/content/10/6/e8454
- Nanozymes (artificial proteins) as desinfectans
https://pubs.acs.org/doi/pdf/10.1021/acsanm.8b00153
- Applications of spider web polymers
https://onlinelibrary.wiley.com/doi/epdf/10.1002/term.2380
- Biosensors using CRISPR/Cas9
https://www.wired.com/story/a‐new‐startup‐wants‐to‐use‐crispr‐to‐diagnose‐disease/
https://synbiobeta.com/mammoth‐biosciences‐launches‐to‐develop‐worlds‐first‐crispr‐based‐detection‐
- Chromosome engineering
https://www.nature.com/articles/d41586-018-05857-9
https://www.nature.com/articles/d41586-018-05309-4
- Diagnostic tool for microbiome
https://www.nature.com/articles/s41467-018-05864-4
- Synbio education kits
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6070313/
http://advances.sciencemag.org/content/4/8/eaat5105
- Biocontainment for algeae
https://www.sciencedaily.com/releases/2018/11/181126123317.htm
- Yeast biohybrids
http://science.sciencemag.org/content/362/6416/813
- CRISPR babies
https://www.nature.com/articles/d41586-018-07559-8

Other
- New method to assess Synbio applications
http://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0168564&type=printable
- Developments in Synbio companies
http://www.sciencedirect.com/science/article/pii/S0167779917300215
http://labiotech.eu/infographic-a-handy-guide-to-the-european-synthetic-biology-startup-scene/
- Safe by design
https://link.springer.com/article/10.1007/s11948-017-9969-0
https://link.springer.com/article/10.1007/s11569-017-0301-x
- IGEM
http://igem.org/Team_Wikis?year=2017
iGEM 2018: http://2018.igem.org/Main_Page  
- Investments in Synbio
https://synbiobeta.com/fifty‐synthetic‐biology‐companies‐raised‐1‐7b‐2017/
http://www.calvinschmidt.info/data/2017/12/30/2017‐review‐of‐synthetic‐biology‐investing


Kind regards,
Boet Glandorf

I am pleased to join the discussion and respond to the disquiet over the definition of “synthetic biology” (Posts #9378, #9372, #9368, #9383). Below I reproduce the AHTEG definition:

“[S]ynthetic 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” (Decision XII/17).

The problem lies in distinct interpretations of words therein, not defined in the CBD or NP.

Whereas broad adjectives (e.g., “new”, “further”, “modern”) will complicate the task of TOPIC 1, distinct interpretations of “material” will preclude any agreement on TOPIC 4 (e.g., does “material” include information or is it just the tangible?). A similar concern would also arise for the word “part” in the definition suggested in Post #9383.  I will elaborate further in the forum next week on TOPIC 4.

Dear participants,
I’m Leyenis García Santos from the Biodiversity and Biosafety Department  belonging to National Biosafety Authority of Cuba. First of all, I would like to congratulate moderators and Secretariat for this on line forum on synthetic biology. I’m please on participate for first time and looking forward to contribute  during exchange of opinions on each topic.
best regards,
Leyenis

Dear colleagues,

My name is Swantje Straßheim and I work at the German Federal Office of Consumer Protection and Food Safety in the secretariat of the German Central Committee for Biological Safety (ZKBS, https://www.zkbs-online.de/ZKBS/EN/02_UeberUns_Aufgaben/ueber_uns_aufgaben_node.html) who carries out a monitoring on Synthetic Biology.

I would like to agree to posts [#9363], [#9364] and [#9381] that there are no new technological developments in synthetic biology since the last meeting of the AHTEG in 2017. In its second report on Synthetic Biology published in July 2018, the ZKBS concluded that were no new technological developments bearing a risk to biological safety.

However, as others have remarked before, there is a great amount of research papers published on Synthetic Biology and existing ideas and techniques have been applied further.
For example, an infectious horsepox virus has been chemically synthesized (Noyce RS, Lederman S & Evans DH (2018). Construction of an infectious horsepox virus vaccine from chemically synthesized DNA fragments. PLoS ONE 13(1): e0188453). This is work can be seen in the context of genome synthesis and builds on research papers where e.g. the complete polio virus genome (Cello et. al. 2002, doi: 10.1126/science.1072266) or the genome of Mycoplasma mycoides (Gibson et al. 2010, doi: 10.1126/science.1190719) were synthesized.
Another example is a biomedical tattoo that detects hypercalcemia associated with cancer. In this approach, a calcium-sensing receptor is rewired with a synthetic signaling cascade that activates the production of melanin as a response to increased blood Calcium (Tastanova et al. 2018, doi: 10.1126/scitranslmed.aap8562). This research is part of broad approaches aiming at constructing biological networks within cells to perform specific tasks, e.g. in the biomedical sector (reviewed in Heallmen & Fussenegger 2015, doi: 10.1016/j.jmb.2015.08.020).

However, I share the concerns put forward in posts [#9372] and [#9382]. The operational definition of Synthetic Biology allows to classify most developments made through modern biotechnology as Synthetic Biology. Most of these organisms are LMOs as defined in the Cartagena Protocol and can be covered by its provisions. 

Best regards,
Swantje

The AHTEG definition of synbio is so broad it could apply to almost any biotech including GMO, transgenics, syngenics, RNA interference, gene editing etc . Synbio undoubtedly is composed of many of these techniques. However I prefer the definition produced at the COP in Korea in 2014 which specifies the types of techniques and outcomes  in Synbio that separate it from other types of biotech ( Updated report UNEP/CBD/COP/12/20 28 August 2014) :
"8. Synthetic biology falls within the scope of biotechnology, as defined by the Convention on Biological Diversity i.e. “... any technological application that uses biological systems, living organisms, or derivatives thereof, to make or modify products or processes for specific use.” Synthetic biology methodologies and techniques share various degrees of overlap with those of “modern biotechnology” and, in particular, the “application of in vitro nucleic acid techniques […] that overcome natural physiological reproductive or recombination barriers and that are not techniques used in traditional breeding and selection” as defined in the Cartagena Protocol on Biosafety.
9. While there is no internationally agreed definition of “synthetic biology”, key features of synthetic biology include the “de novo” synthesis of genetic material and an engineering-based approach to develop components, organisms and products. Synthetic biology builds on modern biotechnology methodologies and techniques such as high throughput DNA technologies and bioinformatics. There is general agreement that the processes of synthetic biology aim to exercise control in the design, characterization and construction of biological parts, devices and systems to create more predictable biological systems. The areas of research that are considered “synthetic biology” include DNA-based circuits, synthetic metabolic pathway engineering, synthetic genomics, protocell construction, and xenobiology: "
Thus this report identified important features which separate synbio from other biotech so perhaps we should focus on advances in the development of de novo systems using these techniques rather than new advances in GM, GEd etc..

Greetings everyone.
My name is Todd Kuiken and I work in the Genetic Engineering & Society Center at North Carolina State University.  I’ve had the honor to be a member of the AHTEG in 2015 and 2017. I would like to offer my congratulations to the moderators and the robust discussions we are already having on this topic.

The definition the CBD has developed around synthetic biology, while important, has always been problematic in setting boundaries about what is in and what is out.  As evidence by the expanding tasks of the AHTEGs and open-online forums, which now incorporate gene drives and genome editing.

Previous interventions in this online forum have raised important points about distinguishing between technology developments since the last AHTEG and applications of those technologies. For instance, the recent publication of using CRISPR technologies to deliver genetic changes to crops through pollen is a new application, but not necessarily a new technology development since the last AHTEG. See:

https://www.sciencemag.org/news/2019/03/corn-and-other-important-crops-can-now-be-gene-edited-pollen-carrying-crispr

As Ms. Margret Engelhard (#9380) and Ms. Boet Glandorf (#9385) have discussed these technologies are potentially influencing many application areas.  Particularly the conservation community.  The International Union for Conservation of Nature (IUCN) will be releasing its assessment of synthetic biology and gene drives on conservation soon (probably March or April) and could be a useful resource for examining the impacts of synthetic biology on conservation as well as new developments in the field.  Full disclosure, I was an author on that assessment.  

What this topic is revealing to me is that the horizon scanning activities that will be discussed in Topic 2 are incredibly important for the CBD to keep pace with the applications these technologies may influence, and the subsequent risk assessment methodologies that may need to be developed.

I look forward to the continued discussion.

Kindest regards,
Todd Kuiken

Regards,

Many thanks to the secretary and the moderators for opening this discussion space in order to consolidate positions based on science about the advances of synthetic biology and how these are framed within the objectives of the agreement.

I hope to contribute positively to this discussion and also learn a lot from everyone.

Thank you,

Carlos Ospina
Ministry of Environment and Sustainable Development
Colombia

Hello everybody,

I'm Emmanuel González-Ortega, from Mexico. Just took office at CIBIOGEM, the Inter Secretarial Commission for Biosafety of Genetically Modified Organisms of Mexico.

I'll be reading and participating in the discussions of this important forum.

Cheers.

Emmanuel

Dear Forum Participants,

I would like to thank you all for your contributions that are enriching our discussion about new technological developments in synthetic biology since the last meeting of the Ad Hoc Technical Expert Group.

Some of you have highlighted that there have not necessarily been material advances in synthetic biology technologies since the 2017 AHTEG report. Nevertheless, many of you have contributed with diverse examples of the applications of different synthetic biology methodologies.

As several have mentioned, the definition of synthetic biology could be a limitation, but considering that our mandate in this forum is to share examples of new developments, I would kindly ask the participants to focus on this forum’s topic, and I’m sure that at the end of this week we will have a good update of the advances made in this field after 2017. Our inputs will be of great benefit for the upcoming meeting of the AHTEG.

I would also like to invite the participants that have not yet commented to post their messages until Sunday.

Kind Regards,

Maria

On the Topic 1 of the Online Forum “New technological developments in synthetic biology since the last meeting of the Ad Hoc Technical Expert Group”.

A recent work published in Science [ http://science.sciencemag.org/content/363/6429/884 ] reports the design and construction of an 8 letter DNA/RNA like molecule (the researchers included 4  additional synthetic nucleotides: P, B, purine like;  Z, S, pyrimidine-like. This molecule is able to mimic the chemical and molecular behavior of the conventional 4-letter DNA molecule: structural properties, information storage, replicative, mutative, evolutive and… apparently, a lot more specific as a translational-messenger molecule (compared to the conventional DNA/RNA molecule).

This landmark work opens a new world of considerations and questions for Biodiversity, Ecology, Evolution and Synthetic Biology; I’d like to mention here a few ones:

What could be the outcomes of the potential presence of organisms produced through synthetic biology containing the 8-letter DNA molecule, once released to the environment, with conventional organisms able to mate with the synthetically produced-LM organisms?

From an epigenetic perspective, how does the complex mechanisms regulating the epigenetic regulation of organisms will behave when facing an 8-letter DNA molecule?

There is yet a large knowledge gap concerning the behavior, regulation and off-target effects of genome editing on conventional 4-letter DNA organisms. The introduction of Hachimoji DNA in organisms could take the complexity of the genome editing systems to an exponential level.

In the Microbial Ecology field; what would be the scenarios in the context of horizontal DNA transmission (i.e. among bacteria)?

In the light of a potential universe (maybe not that much) of new proteins product of the “Hachimoji DNA/RNA" molecules, what could be the scenarios for the field of medicine, specifically, immunology (meaning, new potential allergens)?

Dear participants,

My name is Luciana Ambrozevicius and I work in the brazilian Ministry of Agriculture. First I would like to send my congratulations to the moderators and to the forum participants for all the interesting messages and articles already shared.

I would like to agree to many other posts that there are no new technological developments in synthetic biology since the last meeting of the AHTEG in 2017.  There are many applications of the modern biotechnology, including synthetic biology, most of them in the early stages of research.

Best regards,

Luciana.

Dear Colleagues,

When I mentioned there is no real new technology developed since 2017, I did not mean that all the diverse and novel applications of the technology do not need risk assessment. I completely agree to our colleagues that we shall take care all the newly trend of the application, e.g post [9313] [9399] and their potential uses. Especially there are real cases of novel DNA sequences that have been created. During an interesting conversation, one reputed synthetic biologist predicted that complete new life created from scratch may come to the world in less than ten years.

Hope this can clarify the idea behind my last post.

Best wishes

Wei

Dear all,

Relevant to Topic 1, a review paper published in January 2018 furnishes a helpful outline of recent progress in engineered gene drives and cites a number of potential applications relevant to the modification of sexually reproducing organisms. [Paper: Marshall JM, Akbari OS (2018) Can CRISPR-based gene drive be confined in the wild? A question for molecular and population biology. ACS Chemical Biology 13: 424-430]

Specifically, the paper discusses the use of CRISPR as a gene editing tool and its utility for improving current gene drive engineering technology. The implications of this tool are assessed according to two goals: (1) gene drive systems should be capable of spreading to the extent necessary to achieve the desired impact and (2) gene drive systems should be able to be recalled if needed. 

In particular, this paper notes the relevance of CRISPR for three categories of gene drive systems: (1) “conventional”, or threshold-independent drives that are capable of spreading from a very small initial release, (2) “threshold-dependent”, or drives that must surpass a certain level in the population to persist, and (3) “self-limiting”, or drives that are designed to be eliminated over time.

By way of example: in the first category – “conventional” or threshold-independent drives – CRISPR tools are being investigated for their high level of accuracy when targeting specific sites on a genome in several species. An additional benefit is that this technology could potentially be applicable to removing gene drive systems from the environment in the event of unwanted consequences (i.e. “homing-based drive remediation”).

In the second category - “threshold-dependent” drives - CRISPR technology has recently been employed to engineer “site-specific chromosomal translocations,” which tend to spread into a population if initially present in a majority of organisms, and to be eliminated otherwise. As the paper states, such systems have recently been engineered in several species, “suggesting it is only a matter of time until CRISPR-based translocation drives [are] widely available.”

In the third category, CRISPR technology may be able to accelerate the development of certain types of temporally “self-limiting” drives. In one variety, called “killer-rescue”, CRISPR could assist the development of a toxin-antidote mechanism that is expected to help contain the spatial spread of gene drive. In another variety, called “split drive”, unlinked components of a drive system dissociate and eventually fall out of the population after displaying transient spread.

Regards,

Valeri Vasquez

Dear Colleagues,

I’m Chang Jiang and I work at the Chinese Research Academy of Environmental Sciences (CRAES). It’s my honour to participate the online forum. Firstly, I would like to congratulate the moderators.
I agree with the Weiwei and some colleagues that I don’t see new technological developments in synthetic biology since the last meeting of the AHTEG in 2017. Although there are some new applications of the modern biotechnologies, most of them are in the early stage of researches, and there are still many technical problems to be solved.

Regards

I am Paul Freemont and I am a Professor at Imperial College London. I had the great honour of being part of the AHTEG in 2015 and 2017 and I thank the moderators for this excellent forum and exchange of ideas.

I have been involved in the development of the field synthetic biology in the UK and globally since 2004, I would like to share with colleagues some personal thoughts and also some updates on new technology enablers and I apologise for the overlong post.

I think to revisit the definition of synthetic biology will distract from the main topics of the forum. As many colleagues have stated there are converging definitions globally and at the core of synthetic biology is the aim to designing predictable biological systems at the genetic level including genome editing to whole genome synthesis. Another research area which is developing in the US EU and Asia  is aimed at building living systems from scratch using chemical biology and nanotechnology tools (e.g. see http://buildacell.io/). Related to this is the growing field of xenobiology as exemplified by recent work published in Science ( http://science.sciencemag.org/content/363/6429/884 ) with 8 letter DNA/RNA like molecules. The aim of this field is to explore the fundamental nature of the genetic code with potential developments of new genetic code perhaps exemplified by Floyd Romsberg’s group (https://www.scripps.edu/romesberg/files/Zhang-PNAS-2017.pdf). The concept of an orthogonal central dogma to molecular biology is a very active research area and is discussed here (https://www.nature.com/articles/nchembio.2554). It is likely that at some future point (20 to 100 years) that new organisms or life-like organisms will be constructed which as pointed out by very nicely by Margaret [9380] opens up the debate as to what is  natural and what is man-made although this debate could need a completely new on line  topic and would in my opinion distract from the main focus which is on new technology developments in synthetic biology and how they apply to the CBD.

In term of main stream synthetic biology research globally, uses existing model organisms (single cell prokaryotes, eukaroytes including mammalian cell lines and plant systems)  with aims as diverse as fundamental mechanistic understanding to probiotic therapies to industrial biotechnology. One core concept is the synthetic biology design cycle (Design, Build, Test, Learn) as discussed recently here (https://www.nature.com/articles/s42003-018-0076-9). There are many reviews and government reports on synthetic biology and I believe that this forum and previous ones have listed those citations – if not many are listed here (https://data.plantsci.cam.ac.uk/Haseloff/reports/index.html)

Returning to the Topic in hand, as can be seen from the excellent posts from Nikolay and others [9366; 9370; 9383; 9385; 9393] there has been an acceleration in synthetic biology research globally and the field is maturing rapidly. I uploaded some papers on cell-free synthetic biology to the forum as this had been missed and is a very rapidly growing area where cell extracts are used to transcribe and translated genetic code for specific purposes (see uploaded papers for extensive reviews). Whilst this may not seem completely relevant, the extracts are being made from many different organisms and the genetic code is designed from multiple sources, both of which I think is of interest to the CBD community as are the potential applications (e.g. environmental sensing).

As to whether there are any new technologies since 2017 this could be debatable but the pace of development of tools, processes and protocols is rapid and as such these can be considered as technology and application enablers and I agree with Claudia [9383] that a focus on new applications with direct implications for the CBD would be very useful. One technology enablement area which is rapidly developing globally, is the concept of the bio-foundry where automaton, software, machine learning and metrology is being applied to the synthetic biology design cycle (see http://www.synbicite.com/news-events/2018/jun/25/global-biofoundry-meeting-london-june-2018/)

In terms of horizon scanning, I am very impressed with the quality and thoroughness of the online forums and submitted references/ information sources. I would suggest keeping an open channel for forum members to post relevant developments as they happen which would allow the secretariat and the CBD community and online repository of current knowledge which could be archived into different heading in relation to the convention. The difficulty perhaps is in interpreting this vast and growing research area and knowledge base (particularly by non-experts) in relation to the aims of the convention, which of course is the role of the AHTEG and others – however this brings me to my final point.

As we know research often leads to technology translation which often leads to new start-up companies who aim to apply this technology for specific applications. I would like to suggest that we also scan the new start-up activities in synthetic biology as exemplified by several meetings globally but particularly the synbiobeta meeting in San Francisco which brings together many of the key companies and start-ups in synthetic biology (https://2019.synbiobeta.com/).

I apologies for the overlong response but very much look forward to the further exchange  of ideas in the online forum and thanks again to the moderators for their excellent  guidance.

Dear Colleagues,
I would agree with the previous contributors that most of the progress last year is result of new applications of already existing technologies. Despite of that though there are at least four developments that may require attention in short and middle term and I will try to explain why:

1. Development of synthetic virus like assemblies
In 2018 it was demonstrated that it is possible to design protein that is capable to encapsulate its own genome, a key characteristic of natural viruses (Evolution of a designed protein assembly encapsulating its own RNA genome, Butterfield et al., Nature 552 (7685), 415-420). At the same time was demonstrated that it is also possible to design protein molecules that specifically bind to cell receptors and activate them (De novo design of potent and selective mimics of IL-2 and IL-15, Silva et al., Nature 565 (7738), 186-191). Coupling those two developments should be feasible and will mean that it is we can develop virus like assemblies that can bind their own genomes, bind specifically to cells and get internalised. It will be relatively trivial, though technically demanding to some extend to design a genome that replicates in a specific location inside the cell and in a specific organism. Such systems can be modular, combining different receptor specificities with different genome specificities. The genomes can carry different types of cargo, for example the replication can be triggered only if specific loci are present in the genome, e.g. associated cancer in individual patient. It is hard to overestimate the potential of such systems, they can be used to target specific cell populations that carry specific mutation, and thus can be customised to target only the cancer cells in a patient without affecting the healthy ones. Such virus like reagents can be used to spread into the environment and target only specific pests or disease vectors without affecting related species. Such an approach can be even more efficient than the gene drives (both approaches can be combined). At the same time biosafety and biosecurity risks are pretty obvious.

2. Xenobiology
It has been demonstrated that is possible to incorporate two or four additional nucleotides in the genome and maintain them through replication. Such nucleotides can be used to extend the genetic code. At present that increases the safety of such cells and the non-natural nucleotides should be supplied externally. At the same time, if those developments are combined with protein and metabolic engineering approaches that are used to design in vivo synthetic pathways of those non-natural nucleotides the situation can change radically from environmental point of view (and many others). Such a development seems to be feasible in near or mid-future.

3. Improved metabolism
It is already possible to design novel metabolic pathways or enzymes that give advantages into environment of the organisms that carry them (e.g. Synthetic glycolate metabolism pathways stimulate crop growth and productivity in the field, South et al., Science 363 (6422), eaat9077). One can imagine for example that new improved versions of RuBisCo can be developed or even entirely novel enzymes that catalyse the same reaction. If such metabolic pathways or enzymes are transferred to wild relatives, it is hard to predict what will be the consequences. Just because no such enzymes or pathways have not evolved naturally does not mean that they won’t confer advantages. After all, evolution does not necessary “find” the best solutions; it “finds” good enough solutions.

4. New approaches for domestication
Domestication of wild plants that have not been accessible now seems to be possible within reasonable time scale (Domestication of wild tomato is accelerated by genome editing, Li et al., Nature Biotechnology 36, 1160-1163; De novo domestication of wild tomato using genome editing, Zsogon et al., Nature Biotechnology 36, 1211-1216; Rapid improvement of domestication traits in an orphan crop by genome editing, Lemmon et al., Nature Plants 4, 766-770). While we do not consider that to be synthetic biology, it may have significant consequences both positive and negative for the objectives of the Convention and its Protocols, including for the benefits sharing.

I hope I have been able to show that there were developments last year that deserve attention of the online forum and the AHTEG.

Best Regards,
Nikolay

Greetings Participants,

My name is Dr. Benson M. Kinyagia and I work for the National Commission for Science Technology and Innovation, Kenya as a Principal Science Analyst.  I’ve had the honor to be a member of the AHTEG in 2015 and 2017. I would like to offer my congratulations to the CBD Secretariat, moderators and the robust discussants.
I would like to agree to posts [#9363], [#9364], [#9381] [#9393] and [#9398] that there are no new material technological developments in synthetic biology since the last meeting of the AHTEG in 2017. However, as mentioned by [#9399] new frontiers are coming up like the 8 letter DNA/RNA that don’t exist in nature and their impact on environment in case of release need to be adequately considered. Rapid growth of research and development of new tools as posted by [#9409] their output and their relationship to the objectives of the CBD need consideration. Real cases of novel DNA sequences as posted by [#9406] also need requisite consideration and their potential impact on the natural environment.

Dear All,

Thanks to the Secretariat for this new open - ended forum and Congratulations to both moderators. I am pleased to participate and looking forward for fruitful deliberation.


Best regards.

Dear participants,

I am Filemon Shindume, working for the Ministry of Agriculture in Namibia. I had the great honour of being part of the AHTEG in 2015 and 2017 and I thank and congratulate the moderators for this excellent forum as well as the useful sources and exchange of ideas provided.

Would like to concur with the view that revisiting the definition of synthetic biology will distract us from the main topics of the forum by Paul [#9409].  Would further agree with the previous contributors that most of the progresses since last AHTEG of 2017 are more a result of new applications approach of already existing technologies but the pace of development of tools, processes and protocols is rapid and dynamic with concrete examples as published and shared on this forum.

Best regards,

Filemon N Shindume

Greetings to all dear colleagues,
My gratitude goes out to the moderators. I look forward to participating in this in-depth discussion.

I would first like to echo comments made previously highlighting some of the latest developments, including #9399 on the construction of 8- letter DNA/RNA molecules, and #9380 on the growing number of applications being envisaged and developed for conservation applications.

1) With regards to genome editing conservation applications, I would like to draw attention to the recent publication suggesting the engineering of new adaptive traits, introducing ‘barcodes’ for tracking population dynamics, and somatic gene editing to overcome specific selection pressures [Phelps et al., (2019). Transforming ecology and conservation biology through genome editing. Conserv Biol. doi: 10.1111/cobi.13292.] Such applications raise concerns for biological diversity e.g. gene flow to related species, the prospering of a genome-edited organism at the cost of other important species, unintended molecular effects that could generate novel genotypes/phenotypes. 

Concrete applications of genome editing made recently include the expansion of base editor applications to many organisms including rice, wheat, maize, tomatoes, mice, rats, zebrafish, silkworms and human embryos. A review of the process was recently published [Ranzau et al., (2019). Genome, Epigenome, and Transcriptome Editing via Chemical Modification of Nucleobases in Living Cells, Biochemistry 2019, 58, 330−335]. Concerns remain regarding observed unintended effects at the molecular level including off-target mutations.

2) Other developments have been made in the delivery of genome editing tools such as pollen mediated delivery using a combined haploid-induction/genome editing technique for commercial crop varieties, including wheat and maize [Kelliher et al., 2019. One-step genome editing of elite crop germplasm during haploid induction. Nat Biotech 37].

The use of viruses, as raised by #9399 to deliver transgenes/genome editing tools, or the use of synthetic viruses to induce crop trait alterations is also being developed, see [Pasin F., Menzel W. and Daros J.-A. (2019) Harnessed viruses in the age of metagenomics and synthetic biology: an update on infectious clone assembly and biotechnologies of plant viruses. Plant Biotechnol. J., https://doi.org/10.1111/pbi.13084.c].

These above developments, along with gene drives, are shifting the laboratory to the field, and thus warrant special attention due increased uncertainties and risks with regards to potential spread, controllability and exposure.

3) With regards to the guiding question on progress of gene drive research, there have been some recent publications as already highlighted previously, including the development of new Anopheles mosquitoes that target a highly conserved sequence with the aim of circumventing the major issues of resistance development (Kyrou et al., 2018). Such developments have implications for biological diversity by increasing the chance of any outcrossing becoming established in non-target organisms. Developments in the area of localised drives or molecular remediation strategies are largely at the theoretical stage, though a split-drive system was recently demonstrated in flies [Champer et al. (2019) Molecular safeguarding of CRISPR gene drive experiments].

New suggested gene drive applications include deployment for combatting fall army worm and Bactrocera dorsalis pests in sub-Saharan Africa [Ogaugwu et al., (2019) CRISPR in Sub- Saharan Africa: Applications and Education. Trends in Biotechnology, Vol. 37, No. 3].

On a broader note, the targeting of increasing numbers of pests raises further questions as to who defines a “pest” and in relation to which economic interests and ecological systems, with potential impacts with regards the important ecological roles of certain species/ “pests” in local economies. 

Finally, the transition towards modifying wild populations instead of cultivated species (both with gene drives and genome editing conservation applications) raises added complexities and novel concerns regarding the potential for adverse consequences on semi-natural and natural ecosystems that now go beyond agroecosystems.

I look forward to discussing in more detail, the potential impacts of new developments and applications vis-à-vis the three objectives of the convention, in the later sections of the forum.

Dear all,
I am Jussi Jäntti from VTT Technical Research Centre of Finland where I am a research team leader for Production host engineering.

A lot of highly useful information has already been provided by comments in this forum.

I would like to continue on the theme that Paul Freemont [#9409] brought up.

The engineering biology/synthetic biology field is indeed moving towards increasingly automated high throughput systems for strain engineering. The concept of a design, build, test, learn-cycle is already in use in molecular foundries in the USA (e.g. http://web.mit.edu/foundry/; http://www.londondnafoundry.co.uk/foundry). This approach is not restricted to these centers, but is already part of business operation of several companies (e.g. Ginkgo Bioworks, Amyris, Zymergen).

In addition to the significant UK efforts in this discipline, the EU funded IBISBA project (https://www.ibisba.eu/) is on the ESFRI roadmap, and will expand the availability of strain engineering in Europe.

From biotechnological point of view only a minimal fraction of microbes (species diversity) are currently used that may be suitable for production of the increasingly broad range of chemical compounds in an industrial setting. The development of tools such as CRISPR and high throughput methods for strain variant generation are likely to speed up the use of biological approaches to tackle many of the grand challenges we are facing.

Hello everyone. My name is Jenna Shinen and I am a Senior Advisor contracted in the Office of Conservation and Water, at the U.S. Department of State.  I also served on the 2017 AHTEG on Synthetic Biology.  I am pleased to join the discussion on the forum, and thank the moderators for their work and the thoughtful comments of the other participants in the forum.

I agree with previous posts (#9379, #9383, #9395, and others) that have emphasized new advances in applications rather than on the technologies themselves. Since the 2017 AHTEG, we have seen an increasing number of potentially beneficial applications of biotechnology tools with relevance to the objectives of the Convention.  There is a broad range of new applications and products using synthetic biology technologies such as genome editing, including public health and medicine, agriculture and livestock, industrial uses, species conservation, environmental remediation, water resources management, and invasive species control, to name a few.  Many of these applications have already been helpfully highlighted in previous posts. 

For all of these new products, the United States applies a coordinated, risk-based system to protect the environment and human and animal health, to assess and manage any potential health and environmental risks posed by biotechnology products, and to ensure biotechnology products are safe for the environment, health, research, production, and trade.  This system facilitates oversight of planned introductions of biotechnology products into the environment and focuses on the characteristics of the biotechnology product, the environment into which it will be introduced, and the application of the product – not the process by which the product is developed. 

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

Jenna

Dear all,

my name is Helmut Gaugitsch and I am working at the Environment Agency Austria. I had the honour to participate as a member of the AHTEG on synthetic biology in the previous discussion rounds.

I would like to thank the Secretariat for setting up this important online discussion and our moderators for their valuable guidance.

Like others I would also like to point out that as such no new technological developments have emerged in the field of synthetic biology since the last meeting of the AHTEG. However, within the existing technological range R&D has led to various developments which are worth mentioning and which should be kept into account when observing the field. I would like to mention just a few, as follows:

Genome editing approaches (including highly multiplexed CRISPR-based methods and MAGE) may be used for large-scale engineering of metabolic pathways and networks, see.g. (Chari & Church 2017).

This may also be exploredin plants, however the efficiency of multiplexed editing is still said to be quite low (reports at OECD 2018) and trade-offs vis a vis the frequency for off-target modification activity may impede the feasibility of such approaches.

Nevertheless proof-of-concept studies for multiplexed approaches with different site-directed nucleases were conducted in various crops.  In a recent study in wheat 35 different alpha-gliadin genes out of the 45 genes present in a wildtype line were knocked out using a multiplexed approach (Sanchez-Leon et al., 2018).

Other recent reports indicate that multiplexed genome editing to modify a few target loci only is sufficient to generate phenotypic changes in plants. Two recent publications (Li et al., 2018; Zsögön et al., 2018) indicate the potential of genome editing for an approach called de novo domestication, i.e. to rapidly develop crop lines from wild forms with desired properties like strong resistance towards pathogens or salt tolerance. In both cases relevant agronomic characteristics associated with domesticated tomato plants were established in different lines of Solanum pimpinellifolium by simultaneously editing only 4 or 6 genomic loci, respectively, while maintaining the desired resistance traits present in the wild lines. Among the introduced domestic characteristics were increased fruit number, size, shape and nutrient content of fruits as well as plant architecture and growth characteristics. 

Chari, R., and Church, G.M. (2017). Beyond editing to writing large genomes. Nat Rev Genet 18(12), 749-760. doi: 10.1038/nrg.2017.59.

Li, T., Yang, X., Yu, Y., Si, X., Zhai, X., Zhang, H., et al. (2018). Domestication of wild tomato is accelerated by genome editing. Nature Biotechnology 36, 1160. doi: 10.1038/nbt.4273

OECD (2018). Conference on Genome Editing: Applications in Agriculture [Online]. Paris: Organization for Economic Cooperation and Development (OECD). Available: http://www.oecd.org/environment/genome-editing-agriculture/ [Accessed 20.8.2018].

Sanchez-Leon, S., Gil-Humanes, J., Ozuna, C.V., Gimenez, M.J., Sousa, C., Voytas, D.F., et al. (2018). Low-gluten, nontransgenic wheat engineered with CRISPR/Cas9. Plant Biotechnol J 16(4), 902-910. doi: 10.1111/pbi.12837.

Zsögön, A., Čermák, T., Naves, E.R., Notini, M.M., Edel, K.H., Weinl, S., et al. (2018). De novo domestication of wild tomato using genome editing. Nature Biotechnology 36, 1211. doi: 10.1038/nbt.4272

I am looking forward to further participating in this discussion round and in the following ones over the coming weeks.

All the best

Helmut Gaugitsch

Dear all! My name is Kirsi Törmäkangas and I am working as a gene technology regulator in the Ministry of Social Affairs and Health in Finland. I would like to send my congratulations to our moderators and thank for the possibility to participate in this Forum. I have not participated in online forums before, and I am very impressed in the interesting and informative contributions of the participants. I echo the view of many others that while a multitude of new SynBio applications have been developed since 2017, the technological development has been mainly finetuning. And like some other colleagues, I am still struggling with the broad definitions of SynBio, although I admit that we should not let that prevent the discussions here.

Other participants have already brought up many of the issues I had in mind, but I would like to put more emphasis on the health related applications of SynBio, although several have already have been mentioned. Medical field is quite active, and  quite a few pharmaceutical applications have been developed using SynBio, e.g. for cancer research and targeted anti-cancer drug development, for vaccines (https://phys.org/news/2017-09-synthetic-biology-chlamydia-vaccines.html and http://blogs.nature.com/tradesecrets/2018/01/18/synthetic-vaccines), as well as for diagnostics and toxicity studies(https://pubs.acs.org/doi/full/10.1021/acs.chemrestox.6b00396).

Yet, I am wondering if some of these medical applications actually fulfill the criteria of SynBio, e.g. nanocapsules for drug targeting to relevant tissues (https://www.europeanpharmaceuticalreview.com/news/69901/synthetic-biological-nanocapsule/). Some SynBio applications are organisms or cells, but some are actually cell free systems, such as biosensors (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5609271/), and they may not be able to reproduce outside laboratories. So maybe they are not relevant from the biodiversity point of view of CBD? There are also other SynBio applications where the connection to CBD goals is vague, e.g. synthetic cell models for diseases (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5528198/) or large animals modified for xenotransplantation purposes (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5617878/) or human gene therapy products. For the consideration of SynBio under CBD, it is of course crucial to have a full picture of various fields of applications and developments, but further discussions may be needed as to which ones are the most relevant in the CBD concept.   

As to applications converging biodiversity and health related aspects, I found using SynBio for the monitoring of pathogen evolution quite interesting (https://www.hsph.harvard.edu/news/press-releases/zoonotic-malaria-pathway/).
                
Several colleagues (at least posts #9399, #9406, #9409) referred to applications resulting in altered genomic code  / xenobiology, and the possible effects of this on biodiversity. Adaptive evolution of genomically recoded E. coli has indeed been demonstrated in vitro (https://www.pnas.org/content/115/12/3090h), resulting in a strain more suitable for biotechnology purposes. However, it would be premature to extrapolate fitness changes detected in a laboratory experiment to natural environments which are infinitely more complex.

Dear all,

It is a pleasure to join the Synthetic Biology online forum and share my experience related to the proposed topics. My name is Rodrigo Carvalho de Abreu Lima, a Brazilian lawyer with 14 years of experience at the CBD negotiations and agendas.

Regarding Topic 1, it seems quite relevant to remember that the discussions around SynBio at the CBD started during 2008 with the debates related to “Agricultural biodiversity:  biofuels and biodiversity”. At COP10, in 2010, Parties decided to “apply the precautionary approach in accordance with the Preamble to the Convention, and the Cartagena Protocol, to the introduction and use of living modified organisms for the production of biofuels as well as to the field release of synthetic life, cell, or genome into the environment, acknowledging the entitlement of Parties, in accordance with domestic legislation, to suspend the release of synthetic life, cell, or genome into the environment” (Decision UNEP/CBD/COP/DEC/X/37).

At COP11, Parties adopted the Decision XI/11 - New and emerging issues relating to the conservation and sustainable use of biodiversity – emphasizing the need to seek and compile relevant information about “synthetic biology techniques” on the conservation and sustainable use of biological diversity, especially aiming to understand how and if SynBio would fall within the paragraphs 11 and 12 of Decision IX/29. 

During COP12, in 2014, parties agreed the use of the precautionary approach towards SynBio, in accordance with paragraph 4 of decision XI/11, stressing the role of risk assessment and management when dealing with synthetic biology. Since 2008, Parties did not labeled any technology as synthetic biology, and only in 2016, at COP13, agreed on an operational definition as a starting point to facilitate scientific and technical deliberations, as proposed by the Ad Hoc Technical Expert Group on Synthetic Biology (Decision XIII/17):

“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”.

The work of the ATHEG during 2017 up to COP14, in 2018, as well the online forum, also addressed “recent technological developments” as presented at the ATHEG Report (UNEP/CBD/SYNBIO/AHTEG/2017/1/3). Actually, the Report does not present “recent developments”, but rather, more general statements and arguments related to technologies and possible outcomes. At this stage, it seems quite relevant to mention that Parties did not presented recent developments with positive and/or negative impacts to the objectives of the Convention, leading to a substantive discussion based on fragile evidences about synthetic biology.

It seems critical, when the aim is to address and manage risks to biodiversity, to search for substantial and science based evidences regarding any technology. Assumptions that any technology poses risks against the Convention´s objectives is premature, theoretical and does not based on science. It is worth noting that the precautionary approach underscores risk assessment and management as a core basis, and the approach the discussions took up to now seems to contradict this view.

Synthetic biology encompasses a wide range of technology developments having as a basis the use of recombinant DNA and biotechnology. The definition of biotechnology at the CBD - Article 2: Biotechnology means any technological application that uses biological systems, living organisms, or derivatives thereof, to make or modify products or processes for specific use – could contain all the named “synthetic biology” techniques/products.

Jumping from 2008 to 2016 discussions about SynBio to the last COP14, in 2018, new technologies – gene editing and gene drives – emerged as and were labeled as SynBio with a strong understanding that the work of the Parties should be narrowed towards these technologies.

As the online forum starts to discuss and to gather information about it, it would be extremely useful if concrete and sound evidences are collected to allow a proper understanding. Otherwise, the whole debate will continue to lack scientific grounds, leading Parties to adopt decisions that would not fulfill its objectives, rather, that could create obstacles to innovation. 

I appreciate the opportunity and look forward to having a lively debate about this issue.

Sincerely yours,

Rodrigo C A Lima

Dear participants

I have also previously participated in the AHTEG online forum. I work at a public university.

Adding to Helmut's (#9421, and other) interventions on the issue of applications, and applications I haven't noticed being discussed (apologies if overlooked), is the development of delivery technologies to effect engineering in vivo and in situ. These delivery technologies allow both proteins and nucleic acids, such as genome editing reagents and heritable gene regulators such as dsRNA, to be delivered by topical/inhalation exposure. They can be applied to humans and animals and in some cases insects, nematodes, fungi and plants.

In this intervention, I'll provide some examples of patents rather than focus on the literature. In EP3009511A2 “Novel crispr enzymes and systems” an editing package that includes a CRISPR nucleic acid and cas9 (or similar) protein is envisioned to be delivered to the cells of organisms “via a particle, a nanoparticle, a lipid or a cell penetrating peptide (CPP).”

In patent US9,526,784B2 “DELIVERY SYSTEM FOR FUNCTIONAL NUCLEASES” “Compositions, methods, strategies, kits, and systems for the supercharged protein-mediated delivery of functional effector proteins in to cells in vivo, ex vivo, or in vitro are provided…Functional effect or proteins include, without limitation, transcriptional modulators (e.g., repressors or activators), recombinases, nucleases (e.g.,RNA-programmable nucleases, such as Cas9 proteins; TALE nuclease, and Zinc finger nucleases), deaminases, and other gene modifying/editing enzymes.” “In some embodiments, the step of administering comprises a route of administration selected from the group consisting of oral, intravenous, intramuscular, intra-arterial, Subcutaneous, intraventricular, topical, inhalational, and mucosal delivery.”

In patent US2015/0376586A1 “RNA MODIFICATION TO ENGINEER CAS9 ACTIVITY” “In one aspect, a composition is provided comprising an engineered nucleoprotein complex. In some cases, the engineered nucleoprotein complex comprises a Cas9 polypeptide and a non-natural nucleic acid-targeting nucleic acid, wherein the non-natural nucleic acid-targeting nucleic acid comprises an engineered region selected from the group consisting of an engineered stem loop duplex structure, an engineered bulge region, an engineered hairpin located 3' of the stem loop duplex structure, and any combination thereof.” “Pharmaceutical compositions can be administered in therapeutically-effective amounts as pharmaceutical compositions by various forms and routes including, for example, intravenous, Subcutaneous, intramuscular, oral, rectal, aerosol, parenteral, ophthalmic, pulmonary, transdermal, vaginal, otic, nasal, and topical administration.”

In patent WO2013025670A1 “Methods and compositions for introduction of exogenous dsrna into plant cells” is claimed “a method to silence an endogenous target gene expression in plants by applying a specific dsRNA onto the exterior surface of a plant…The present invention enables the regulation of gene expression in plants. In some embodiments of the invention, the dsRNA is directed to an essential gene of a plant pathogen or pest, whereby the pathogen and/or pest damage is controlled, resulting in desired agronomic performance.” "This invention disclosure is a novel approach of plant hormone-mediated penetration of dsRNA into plant cells and the subsequent induction of plant endogenous gene silencing by application of the dsRNA to a surface of a plant structure, e.g. a leaf surface."

This intervention is not intended to be a comprehensive listing of relevant technological developments of extra-laboratory application of synthetic biology tools, but to simply acknowledge the expanding range of applications, environments in which engineering can or may be done, and the co-technologies, eg delivery, that are advancing as rapidly as the tools of genome modification.

With regards to all
Jack

Dear forum participants,

My name is Felicity Keiper and I work in regulatory policy in the agricultural biotech industry. I have experience in biotech research, risk assessment and management for LMOs for environmental release, and the regulatory requirements for commercialization of agricultural biotech products. I have been following this topic under the CBD, and all topics under the Cartagena Protocol, for several years, and was fortunate to participate in the AHTEG on Synthetic Biology in 2015.

I agree with many points raised by others in this discussion so far: that many of the reported new developments in synthetic biology are new applications of established technologies (#9371, #9364, #9363, #9385, #9393, #9408, #9413, #9418, #9421, #9423); that the technology used or degree of genetic modification does not make something “synthetic biology” (#9376); that the same product can be created using different tools (#9368); and that the most important considerations are the characteristics of the product (organism) (#9368), and the intended application (#9368, #9383, #9408).

The issue of the problematic definition arises in every discussion on synthetic biology under the CBD. I agree with others that it is difficult to determine at what point genetic modification or genome editing becomes “synthetic biology”, or differentiate it from “biotechnology” as defined by the CBD or “modern biotechnology” as defined by the Cartagena Protocol. In my view, it cannot be differentiated, and it’s a moot point: work to date under the CBD (including online discussions, information submissions and AHTEGs in 2015 and 2017, and the information submission of February 2019: https://bch.cbd.int/synbio/submissions/) and the Cartagena Protocol (online discussions in 2016: http://bch.cbd.int/onlineconferences/2014_2016period.shtml) has not identified a current or foreseeable organism developed using “new” technologies that is not within the scope of existing national/regional/international regulatory mechanisms applicable to biotechnology. This is pointed out again here (ZKBS report #9393) and in topic 2 (#9390, #9405).

In the recent information submission by the Global Industry Coalition (February 2019: https://bch.cbd.int/database/record.shtml?documentid=114285), we provide a review of developments in biotechnology (that may be labeled synthetic biology) reported in the scientific literature, many of which have been provided by others in this discussion. While we review applications of genome editing in plants in our submission, this is for the purpose of sharing factual information on the status of technological development and application – we emphasize that we do not consider these developments to be “related to synthetic biology”. Genome editing is a broad category of enabling tools that can be applied to achieve the same outcomes as established technologies, and where the outcome is an LMO/GMO, we are not aware of a case which has challenged existing regulatory processes.

We strongly agree with the comment in #9383 that for these discussions to be useful, there needs to be a focus on new applications, and we add that they must have a realistically foreseeable outcome, be relevant to the scope of the CBD, be outside the scope of existing regulatory mechanisms, and not be duplicating work under the Cartagena Protocol. In regard to the latter, examples of gene drives have been raised in this discussion – organisms containing engineered gene drives are LMOs within the scope of the Cartagena Protocol, and risk assessment for this “new” application is currently under consideration in that forum.

Thank you, colleagues and moderators for a lively discussion on this matter.  I appreciate the summary presented by Mr. Rodrigo C. A. Lima on the earlier discussions and decisions related to synthetic biology, which provides a historical perspective for those of us who were not in on the initial engagements on this topic.  Likewise appreciated are the posts from colleagues that provided a wide range of materials in their reading lists.  However, I think that unless we can clarify the scope of synthetic biology, and among ourselves determine whether or not this suite of techniques fall within CPB's definition of modern biotechnology, most of our discussions will be moot albeit academically interesting.  I have seen previous posts also calling for a clarification on what is included in the term synthetic biology, as operational definitions have a tendency of being left behind by the rapid developments in the various -omics and molecular modification techniques.  As a case in point, do we include developments in epitranscriptomics in our surveys/horizon scanning? and should advances in gene delivery systems, coupled with gene editing techniques be classified as new "developments in synthetic biology"?

Dear forum colleagues

It seems that  the definition of synthetic biology continues to pose a major debating point int he Forum and unfortunately this has existed since the start of the field (late 1990's). To try and clarify,
I thought it might be useful to new forum members or colleagues new to the field to learn about where the name synthetic biology came from - so I have included a quotation from an eminent Polish molecular biologist  and visionary  Waclaw Szybalski see https://en.wikipedia.org/wiki/Wac%C5%82aw_Szybalski

Professor Szybalski stated in a paper in 1974 during the molecular biology technology revolution - "let me now comment on the question "what next". Up to now we are working on the descriptive phase of molecular biology. ... But the real challenge will start when we enter the synthetic biology phase of research in our field. We will then devise new control elements and add these new modules to the existing genomes or build up wholly new genomes. This would be a field with the unlimited expansion potential and hardly any limitations to building "new better control circuits" and .... finally other "synthetic” organisms, like a "new better mouse". ... I am not concerned that we will run out of exciting and novel ideas, ... in the synthetic biology, in general.”

So the term synthetic biology is not knew - however our technological advances particularly in chemical DNA synthesis, are now allowing the realisation  (in part) of  Pofessor Szybalski's vision and the interdisciplinary field of synthetic biology has now emerged which in the main is focused on reprogramming living systems at the genetic level for both specific applications and fundamental understanding. As I mentioned in my other post there are areas of research that want to build from the bottom-up synthetic cell or cell-like 'living' systems and one could argue that these are truly synthetic.

However, my own personal view is that all new molecular biology tools that allow the re-engineering of genetic material or modification of the function of genetic material (including new code)  or complete chemical re-synthesis of genetic material  should be looked at in the context of the CBD. The technologies that implement these approaches (e.g. genetic delivery systems etc, biofoundries etc) should also be included. The definition of what is and what is not synthetic biology is perhaps a moot and academic point which really does not progress the important discussions  that need to be had, in the context of the acceleration of  biotechnology capabilities.

At the last AHTEG, I believe we did discuss a list of developing molecular biology/biotechnology tools which was tabled by the Chair? (sorry can't remember) and was extremely useful to inform the discussions. Perhaps the horizon scanning exercise can formalise this and list technology developments, describe what they are,  how they are implemented and how they are relevant to the 3 aims of the CBD. Such a list could expand and grow and be an incredibly useful resource for the CBD community in its deliberations.

Topic 1: New technological developments in synthetic biology since the last meeting of the Ad Hoc Technical Expert Group


Dear Colleagues

Hello, My name is Jim Thomas - I am Co- Executive Director of ETC Group - a civil society organization that monitors emerging technologies (including emerging biotechnologies) and I had the privilege of also serving on the past 2 AHTEGs on Synthetic Biology.  ETC would like to join with the appreciation expressed to the CBD Secretariat and the moderators  for making this forum available and for the high quality and constructive discussion that is now underway here.

With regard to the questions raised regarding  definition of Synthetic biology we would concur that the operational definition as agreed by the parties to the CBD is indeed a useful starting point for these kind of processes and agree with others that it is not constructive to try to reopen the long definitional debates we have already had here. The broad nature of the definition is particularly useful in horizon scanning exercises such as this one given the rapid change and development of the field. To my mind the term synthetic biology , particularly as it relates to the governance discussions at the CBD, should less be regarded as a technically limited subfield of modern biotechnology whose delineations need to be circumscribed and litigated but more as signifier that the parties and others engaged in this process are attempting to ensure that novel biotechnological methods, applications and approaches (going beyond established  ‘first-generation’/transgenic means) are appropriately monitored, assessed and where necessary brought under oversight. Since the Convention’s interest here  is to understand novel threats, opportunities and complicating factors in achieving the conservation and sustainable use of biodiversity (and equitable sharing of benefits) allowing this forum and other processes to widely scan developments can only bring increased knowledge, light and foresight to the process of good governance and safeguarding biodiversity, cultures and livelihoods. I do appreciate however Professor Paul Freemont’s proposal (#9440) that it could be helpful to develop an indicative list of technologies and approaches to help guide discussion - but such a list should be non exclusive - that is to say open to continued expansion as new techniques and approaches emerge.

I am grateful to many of the excellent suggestions pointing to new technological developments in the field of synthetic biology/new modern biotechnologies. I would gently question whether its necessary to only identify technological development that have strictly come into being since the last AHTEG. Since technological development always emerge on a continuum its also helpful to identify those who have become more significant, prominent or whose relevance to the aims of the convention may not have been so apparent two years ago.

For example ETC Group believes that the Convention should now be paying closer attention to developments in the area of “transient expression” where  crop biomass is tranfected or infiltrated by an engineered bacterium, virus or other carrier such as a nucleic acid-associated nanoparticle in order to initiate production of viral particles, natural product synthesis , gene silencing or similar in a temporary manner that doesn’t stably integrate the novel genetic material into the genome. Transient expression has been under development for some years but is now accelerating at a significant pace. Such  systems include new work on  contained production systems  for high value compounds (for example this paper showing production of a range of terpenoids in tobacco leaf cultures from transfection by engineered agrobacterium https://link.springer.com/article/10.1007/s00299-018-2296-3 - this technology being commercialized as the HyperTrans system  by Leaf Expression Systems of UK (https://www.leafexpressionsystems.co.uk). These contained systems may have implications for the sustainable use of biodiversity since economies of natural product production are closely tied to sustainable use - as we have seen with the case of replacing naturally grown vanilla, stevia, artemisia and other botanical  ingredients by synthetic microbial production). The recent extension of transient expression technology into production of natural products through ‘in planta’ production  is of key interest to the aims of the convention should that move to a commercial stage.

Secondly transient expression systems are now being developed with an eye towards large scale and/or open air use in crops. Recent examples include development of spray methods for transfecting whole plants  or ‘airbrushing’ leaves with engineered agrobacterium to initiate transient expression on the field. A particular focus is on transfecting crops such as tobacco to transiently express cellulases that would break down crop biomass ready for production of ethanol and other bio-based chemicals. Such a system if effective could enable changes in land use, cropping and biomass extraction that could have significant aggregate impact on biodiversity, cultures and livelihoods. Researchers also recently reported on using transient expression to initiate plant protection in leaves against fungus as well as use of nanoparticle mediated transient expression in plants (with siRNA rather than Agrobacterium - see below) for gene silencing in crop plants. The prospect of genetically active engineered ‘transient expression’ vectors as a new form of crop pesticide  should be of particular concern to the CBD and it speaks to the trend expressed also by Eva Sirinathsinghji of Third World Network n her response that we are seeing the location of  genetic engineering and genetic transformation move from ‘in vitro’ to ‘in situ’ - that the ecosystem is becoming the lab.

There is of course a very direct link to the developments that Dr Engelhard raised with regard to the growing interest in use of engineered viruses and also HEGAA’s that has become more pronounced since the last AHTEG. (see http://web.evolbio.mpg.de/HEGAAs/).  While HEGAA’s were raised in passing during the last online forum there seems to be renewed technical attention on horizontal spread of engineered viruses for transforming crops and plants in situ - potentially via insect vectors as in DARPA’s insect allies programme. As best as I understand the aim here also is to induce a form of transient expression by virus infection, Use of engineered  siRNA in RNA sprays is also a transient expression system of sorts, targeted at environmental use that relies upon synthetically engineered nucleic acids. For example Finnish and French  synthetic biology researchers reported last year on spraying synthetic double stranded RNA on barley to act as a fungicide (https://phys.org/news/2016-10-antifungal-rna-barley-crop-disease.html) and Australian researchers earlier reported using nanoclay particles as a spray carrier for dsRNA in a way that gives crop protection for up to 30 days (https://www.nature.com/articles/nplants2016207). use of engineered RNA sprays will not be limited to crops and agriculture. e.g. Forrest Innovations of Israel is applying RNA spray to reverse pyrethroid resistance in mosquitos - http://www.forrestinnovations.com/images/Larval-applocation-of-sodium-channel-Forrest-paper-Parasites--Vectors-2016.pdf)

Another class of synthetic biology innovations that fall into this trend of moving to the field is the application of synthetically engineered “biologicals" (microbes) to agriculture. Particularly urgent for CBD to address is the upcoming commercial release of nitrogen fixing synthetically engineered /gene-edited bacteria developed by PivotBio for corn, soy and Rice production. Pivot claims that its gene edited microbe has been trialled in over 5000 field trial plots and the company says it is scheduled for sale this year. Although Pivot Bio  market their “ProveN” microbe as "non-transgenic”  , the company uses gene-editing tools in their production (see https://www.wired.com/story/farmers-can-now-buy-designer-microbes-to-replace-fertilizer/). Joyn Biotech is another synthetic biology firm formed by a 100 million US  dollar joint venture between Bayer and Ginko Bioworks to deploy synthetically engineered microbes for nitrogen fixation in agricultural soils. Joyn say they are 2-3 years away from deployment. Perhaps those on this forum from Bayer can comment on this timeline.

A number of other respondents have brought up developments in gene drives (e.g. development of mammalian gene drive systems). In 2018 ETC Group released an overview report on the application of gene drive systems to agriculture (See Forcing the Farm: http://www.etcgroup.org/content/forcing-farm) which we hereby submit for consideration of new developments in the field. Our research show that there is  increasing  work on application of gene drive systems to agricultural pests (Especially insects and aphids) as well as to applying gene drive as a breeding tool for livestock. To date we cannot identify successful use of CRISPR gene drive systems in plants (although perhaps others on this forum can correct that) or any working examples of so called local or controllable gene drive systems beyond theoretical models. Given that some of these  theoretical ideas are advanced in policy fora as if they exist I think it may be important for the moderators in their summary of this forum to also point to and clarify  expected developments that in fact have not happened (i.e. local/controllable/limited gene drives). One interesting and concerning technological development that similarly as far as we can tell  has not happened but is already covered by patent protection is work describing use of gene drives to spread  optogenetic traits in insects including honeybees and mosquitos with the intention of thereby controlling insect behaviour using a light beam - see http://www.freepatentsonline.com/y2016/0310754.html. Honey bee experts we have spoken with tell us that this is not a viable technology given the complexity of honey bee olfaction but the fact that Elwha inc has even invested significant time in seeking to obtain this patent is concerning. It would portend a technological system where insect behaviour - including pollination and other ecological services - might be directed by proprietary light beams.

It may also be appropriate to identify as new developments  the rise of work on  storage of Data on synthetic DNA. Recent article in Wired reports that the Defense Advanced Research Projects Agency last year granted $15.3 million in grants to discover new biochemical ways to store binary and that Microsoft plans to have an operational prototype storage system based on DNA working inside one of its data centers by 2020. - https://www.wired.com/story/the-rise-of-dna-data-storage/

I would finally like to suggest that as well as making a list of developments in the field (and Proffessor Freemont's proposed list of techniques./approaches etc) it may be also useful for the moderators to identify the trends that these represent. In ETC Group’s point of view the renewed emphasis on technologies such as biologics/soil microbes, RNA Sprays, , HEGAA’s, engineered insects and gene drives point to a move away from engineering of crops, food etc towards a focus on re-engineering  ecosystems through genetic tools - including both wild and domesticated ecosystems. (moving from GMO’s to GME’s - genetically modified ecosystems).  We think it is a very pertinent question for the CBD to ask whether the rise of synthetic ecosystems (including though genetic alteration) raise questions following from the interaction of different application that cannot be adequately addressed by case-by-case one at a time consideration of the applications which miss the interaction and larger picture of synthetic ecological interventions emerging together . This is relevant to the point raised earlier about use of synthetic biology/modern biotech for conservation applications but also for the shift from synthetic agroecosystem defined by chemical input to synthetic agroecosystems that will be more and more defined by genetic tools.

best

Jim Thomas
ETC Group

Dear all,

my name is Marc F Schetelig and I work as a professor at the Justus-Liebig-University Gießen, Germany leading the Department for Insect Biotechnology in Plant Protection. To keep it short in response to Maria's request to stick to novel technological developments, I agree with several posts from colleagues that there are no new technological developments to synthetic biology.

Many applications of known technologies have been covered in the references from Nikolay [#9366], Boet [#9385], and Valeri [#9407] and I do think that genetic modifications with CRISPR/Cas and comparable technologies can be modular construction systems as well - combining the fields of gene modification and synthetic biology. 

But I want to make a comment to Maria [#9398] on the definition of synthetic biology. I do think that the limitation of the term is, that it includes many different aspects and fields now that widen our 'novel application' list and references and I am not sure that this is the main idea for this forum.
To keep it short and concise: if the term synthetic biology includes GM applications, we could give up the term GM completely and only use synthetic biology, because scientifically, it does not help to create "new fields" that only exist on paper or are made up to fulfill a positive or negative cliche while combining other already defined fields.

Looking at applications especially in the insect field, the additional references below could be of interest to our ongoing discussions.

Thanks and I am looking forward to more interactions with all of you

Marc


-------

References

Abudayyeh, O. O., J. S. Gootenberg, S. Konermann, J. Joung, I. M. Slaymaker, D. B. Cox, S. Shmakov, K. S. Makarova, E. Semenova, L. Minakhin, K. Severinov, A. Regev, E. S. Lander, E. V. Koonin and F. Zhang (2016). "C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector." Science 353(6299): aaf5573.

Aumann, R. A., M. F. Schetelig and I. Hacker (2018). "Highly efficient genome editing by homology-directed repair using Cas9 protein in Ceratitis capitata." Insect Biochem Mol Biol 101: 85-93.

Baltzegar, J., J. C. Barnes, J. E. Elsensohn, N. Gutzmann, M. S. Jones, S. King and J. Sudweeks (2018). "Anticipating complexity in the deployment of gene drive insects in agriculture." Journal of Responsible Innovation 5: S81-S97.

Buchman, A., J. M. Marshall, D. Ostrovski, T. Yang and O. S. Akbari (2018). "Synthetically engineered Medea gene drive system in the worldwide crop pest Drosophila suzukii." Proc Natl Acad Sci U S A 115(18): 4725-4730.

Callaway, E. (2017). "Gene drives thwarted by emergence of resistant organisms." Nature 542(7639): 15.

Champer, J., J. Liu, S. Y. Oh, R. Reeves, A. Luthra, N. Oakes, A. G. Clark and P. W. Messer (2018). "Reducing resistance allele formation in CRISPR gene drive." Proc Natl Acad Sci U S A 115(21): 5522-5527.

Drury, D. W., A. L. Dapper, D. J. Siniard, G. E. Zentner and M. J. Wade (2017). "CRISPR/Cas9 gene drives in genetically variable and nonrandomly mating wild populations." Sci Adv 3(5): e1601910.

Esvelt, K.M. (2017). "Precaution: Open gene drive research". Science 355(6325), 589-590. [doi: 10.1126/science.aal5325]

Hallmann, C. A., M. Sorg, E. Jongejans, H. Siepel, N. Hofland, H. Schwan, W. Stenmans, A. Muller, H. Sumser, T. Horren, D. Goulson and H. de Kroon (2017). "More than 75 percent decline over 27 years in total flying insect biomass in protected areas." PLoS One 12(10): e0185809.

KaramiNejadRanjbar, M., K. N. Eckermann, H. M. M. Ahmed, C. H. Sanchez, S. Dippel, J. M. Marshall and E. A. Wimmer (2018). "Consequences of resistance evolution in a Cas9-based sex conversion-suppression gene drive for insect pest management." Proc Natl Acad Sci U S A 115(24): 6189-6194.

Kyrou, K., A. M. Hammond, R. Galizi, N. Kranjc, A. Burt, A. K. Beaghton, T. Nolan and A. Crisanti (2018). "A CRISPR-Cas9 gene drive targeting doublesex causes complete population suppression in caged Anopheles gambiae mosquitoes." Nat Biotechnol 36(11): 1062-1066.

Li, J. and A. M. Handler (2017). "Temperature-dependent sex-reversal by a transformer-2 gene-edited mutation in the spotted wing drosophila, Drosophila suzukii." Sci Rep 7(1): 12363.

Marshall, J. M., A. Buchman, C. H. Sanchez and O. S. Akbari (2017). "Overcoming evolved resistance to population-suppressing homing-based gene drives." Sci Rep 7(1): 3776.

Nash, A., G. M. Urdaneta, A. K. Beaghton, A. Hoermann, P. A. Papathanos, G. K. Christophides and N. Windbichler (2018). "Integral gene drives for population replacement." Biology Open: bio.037762.

Noble, C., J. Olejarz, K. M. Esvelt, G. M. Church and M. A. Nowak (2017). "Evolutionary dynamics of CRISPR gene drives." Sci Adv 3(4): e1601964.

Prowse, T. A. A., P. Cassey, J. V. Ross, C. Pfitzner, T. A. Wittmann and P. Thomas (2017). "Dodging silver bullets: good CRISPR gene-drive design is critical for eradicating exotic vertebrates." Proc Biol Sci 284(1860).

Reed, F. A. (2017). "CRISPR/Cas9 Gene Drive: Growing Pains for a New Technology." Genetics 205(3): 1037-1039.

Unckless, R. L., A. G. Clark and P. W. Messer (2017). "Evolution of Resistance Against CRISPR/Cas9 Gene Drive." Genetics 205(2): 827-841.

Yan, Y., R. J. Linger and M. J. Scott (2017). "Building early-larval sexing systems for genetic control of the Australian sheep blow fly Lucilia cuprina using two constitutive promoters." Sci Rep 7(1): 2538.

Just to extend the discussion on delivery methods of new GM/synbio techniques that are transferring the laboratory to the field, and the issue of spray methods raised by comment #9442, I would like to draw attention to a recent publication in the therapeutic field working on the development of aerosol sprays to deliver genome editing machinery. [Zhang H, Bahamondez-Canas TF, Zhang Y, Leal J, Smyth HDC. (2018). PEGylated Chitosan for Nonviral Aerosol and Mucosal Delivery of the CRISPR/Cas9 System in Vitro. Mol Pharm Vol 15(11):4814-4826].

Thank you

Dear all,

I would like to start by thanking the Secretariat and our moderators Casper Linnestad and Maria de Lourdes Torres for this forum. In addition, to thank the participants for this interesting exchange.  It’s a pleasure to participate.

It is nice to see that many people share a similar opinion on topic 1. I echo the view of many that research continues, there were plenty of publications on refinements on tools and extension of ideas into new applications, but I did not see any new technological developments since the last meeting of the AHTEG in 2017 (as already stated in many posts, like #9363, #9364, #9371, #9379, #9381, #9393, #9404, #9408, etc.

The term Synthetic Biology was used already a long time ago, see the title of the book from Leduc in 1912 (Leduc S. La biologie synthétique. Paris: A. Poinat; 1912). The enabling tools advances and the opportunities the field offers contributed to great public investments and astonishing progress in research mostly over the past two decades or so. But finding a consensus definition for what is the concept of synthetic biology has proven to be challenging.  Most times whether you call it synthetic biology or LMO/GMO will be both correct because it makes no difference whether there was "de novo”  synthesis or not of the inserted DNA (also in line with the conclusions by AHTEG reports from 2015 and 2017).  I agree with those (such as #9409, #9383, etc.) that though there is no agreement on the definitions, there are significant similarities. One point I believe it is very important to raise is that genome editing is a range of tools/technologies we can use in multiple ways. We can use to insert, replace or remove. We can make from a single base mutation (similar to those common in nature) considered by some countries as conventional breeding for regulatory purposes. To the development of an LMO. For genome editing, its mere use does not mean it is synthetic biology. It is the product/organism obtained, not any in the group of technologies within genome editing that can tell if it is considered and regulated as LMO/GMO or conventional breeding.

best regards, Lúcia

Dear Casper and Maria
I think that no new techniques have emerged since the last AHTEG meeting. Yet there are plenty of achievements in the application of those established techniques and many of them require new or modified risk assessment techniques other than those already established.

However, going through the AHTEG report,(and please rectify me if I am wrong) I did not find any list of those techniques. Even in the introductory part of this forum, I found no clue of the existing technological developments in synthetic biology during the last meeting of the Ad Hoc Technical Expert Group to see if something is missing.

In the upcoming few lines I will try to list some of those techniques and concepts:
Clustered Regularly Interspersed Short Palindomic Repeats (CRISPR), Directed Evolution, DNA-based genetic circuits, DNA Synthesis and Assembly,
Epigenetic Modification, Expanded Genetic Alphabets, Genome Editing, Genome-level Engineering, Genome Shuffling, Gibson Assembly, Minimal Genomes,
Multiplex Automated Genome Engineering, Oligonucleotide Directed Mutagenesis, Protocell Construction, Refactoring of Genomes, RNA-Directed DNA Methylation (RDDM), RNAi (RNA Interference), Standard Modular DNA ‘parts’ or ‘Biobricks’, Synthetic Metabolic Pathway Engineering, Synthetic Genomics, Transcription-Activator-like Effector Nucleases (TALENs), Xenobiology, Zinc Finger Nucleases(ZFN)..

With regards to the definition of Synthetic biology, I believe that the SynBio AHTEG already proposed a definition and it was adopted by the COP Decision XII/17. Openning the discussion on Definition right now will just shift the forum away from it intended purpose,

Warm regards,
Prof.Dr. O.A.EL-Kawy

Dear Participants
I am Geoff Turner, currently working at Imperial College London, and have spent many years supporting regulatory matters across a breadth of areas in both government and the private sector, and now the University sector.
In referring to the December 2017 report of the AHTEG (CBD/SYNBIO/AHTEG/2017/1/3), the list under article 3.1 para 15 a) - k) provides a snapshot of what was considered a "recent technological developments…in synthetic biology" at that time.  The listing presents a range of applications, many of which clearly fall under the definition of LMO under the Cartagena protocol, and as such it seems appropriate they are managed under legal frameworks and guidance derived as implementations of the Protocol in member states.  The scope of technological applications which should be considered under the theme of "Synthetic Biology", current working definitions aside, should thus be those which clearly fall outside of the scope of the Protocol.  Many technological developments which are not LMOs under the Protocol have been cited in this thread, and I imagine the pace of technological change will ensure a steady pipeline in the foreseeable future
Within that basket of "new technological developments", the question as stated still poses a challenge in that technology development in itself is an iterative process whereby incremental gains made in discovery phases today, will inform further discovery tomorrow. A developmental pathway may have been derived from a theory moving to proof of concept,  or a tangent derived from an entirely different research direction.  Technology development is generally an evolutionary process built upon an ever changing state of knowledge and limited by the range of available tools at the time.  Revolutionary breakthroughs do occur, and certain gene editing tools might fall into that category, thereby enhancing the molecular toolbox for technology developers.  The question for this forum is at what stage of technological maturity are we to consider a particular  development a "new technological development" considering that some ideas may never come to fruition if they fall flat anywhere along the developmental pathway.  If this activity is to take stock of "new technological developments" in synthetic biology since the last meeting of the Ad Hoc Technical Expert Group (AHTEG) in 2017, an objective assessment of the technology readiness of proposed developments should be undertaken under a defined set of criteria,  or efficiencies will be lost  in sifting through an ever mounting pile of early phase discovery, much of which may never materialise.  The use of technology readiness levels scales in public sector innovation has been recently reviewed (https://www.innovation.cc/discussion-papers/2017_22_2_3_heder_nasa-to-eu-trl-scale.pdf), and may stimulate thinking as to how to prioritise efforts of the AHTEG in examining the ever increasing breadth of technological developments. 
Objective assessment criteria for technological maturity, coupled with the recognition that applications which are considered LMOs already fall under Cartagena Protocol and can be dealt with as such,  should help to focus efforts on emerging applications of synthetic biology most relevent to the objectives of the Convention.
Thank you for the opportunity to comment

Topic 1:
Greetings to all. My name is Nada Babiker Hamza, Director. Commission for Biotechnology & Genetic Engineering, Sudan.  It is my first time to participate in online forums on synthetic biology, I am so pleased by the discussion and the wealth of information given by the participants. Thanks to the Secretariat and the moderators for giving me this opportunity.
I would like to agree to posts [#9363], [#9364] and [#9381] and others that there are no new technological developments in synthetic biology since the last meeting of the AHTEG in 2017. However, as others have remarked before, there is a great amount of research papers published on Synthetic Biology and techniques have been applied further.
Mr. Jeremy (#9378) mentioned the definition of Synthetic biology of the Royal Society: "Synthetic biology is an emerging area of research that can broadly be described as the design and construction of novel artificial biological pathways, organisms or devices, or the redesign of existing natural biological systems." I find this definition reliable to cover existing technologies including biotechnology.
The excellent interventions reflected a tremendous amount of published research papers in prestigious journals on Synthetic Biology and several innovative techniques applied on different organisms. I agree with Claudia (#9383) and Mr. Paul (#9409) to only consider the new applications with direct implications for the CBD.
Mr. Wei Wei (#9406) mentioned a scientist predicted that complete new life created from scratch is likely to exist in less than ten years.  This was assured by Mr Paul (#9409) predicting it to happen in 20 to 100 years. Are we ready for such life changing events? It is high time to consider with these rapid developments, the risk assessment procedures and methods, they should be updated and adapted with the same pace of the development. Also, their socio-economic impact indigenous people and local communities. I also have the same considerations as (#9399) about the reliability of the possible impacts of the introduction of Hachimoji DNA in organism into nature, the potential effects to the wild types and the biodiversity as a whole.

Hello, my name is Christoph Then, working for the non-profit organisation of Testbiotech (http://www.testbiotech.org). Within the AHTEG I am representing ENSSER, the European Network of Scientists for Social and Environmental Responsibility. There are several new developments which should be discussed by the expert group. Please see some examples below, I grouped them into categories to ease further discussion.

> There are several publications showing that genome editing can induce far reaching changes in organisms, even if no additional genes get inserted. For example:

Sanchez-Leon S., Gil-Humanes J., Ozuna C.V. Gimenez M.J, Sousa C., Voytas D.F., Barro F. (2018) Low-gluten, nontransgenic wheat engineered with CRISPR/Cas9, Plant Biotechnology Journal (2018) 16, pp. 902–910, doi: 10.1111/pbi.12837

Zsögön A., Čermák T., Naves E.R., Notini M.M. Edel K.H., Weinl S., Freschi L., Voytas D.F., Kudla J., Pereira Peres L.E. (2018) De novo domestication of wild tomato using genome editing, Nature Biotechnology 36, 1211-1216

> There are new developments involving insects and their interaction with microorganisms or virus:

Leonard, S. P., Perutka, J., Powell, J. E., Geng, P., Richhart, D. D., Byrom, M., Barrick, J. E. (2018). Genetic Engineering of Bee Gut Microbiome Bacteria with a Toolkit for Modular Assembly of Broad-Host-Range Plasmids. ACS Synth Biol, 7(5), 1279-1290. doi: 10.1021/acssynbio.7b00399

Reeves, R. G., Voeneky, S., Caetano-Anolles, D., Beck, F., & Boete, C. (2018). Agricultural research, or a new bioweapon system? Science, 362(6410), 35-37. doi: 10.1126/science.aat7664

> many articles show that new methods are extending to technical applications beyond what was feasible in 2017:

Buchman A, Marshall JM, Ostrovski D, Yang T, Akbari, OS (2018) Synthetically engineered Medea gene drive system in the worldwide crop pest Drosophila suzukii. Proc Natl Acad Sci 2017:13139. https://doi.org/10.1073/pnas.1713139115

Grunwald HA, Gantz VM, Poplawski G, Xiang-Ru, SX, Bier, E., & Cooper, K. L. (2019) Super-Mendelian inheritance mediated by CRISPR–Cas9 in the female mouse germline. Nature 566:105-https://doi.org/10.1038/s41586-019-0875-2

Hoshika S. et al., (2019) Hachimoji DNA and RNA: A genetic system with eight building blocks, Science, 363:884–887, DOI: 10.1126/science.aat0971

Shao, Y., Lu, N., Wu, Z., Cai, C., Wang, S., Zhang, L. L., Qin, Z. (2018). Creating a functional single-chromosome yeast. Nature, 560(7718), 331-335. doi: 10.1038/s41586-018-0382-x

Dear all,
Thanks for the inclusion of the references about the SynBio technological developments that have been presented here. It is clear that several applications are being developed, although no new breaking technique has appeared in the last two years. Although the information presented here is very valuable, I think is very important to focus on what par. 3 of Dec. 14/19 says: “reviewing new information regarding the potential positive and potential negative impacts of synthetic biology vis-à-vis the three objectives of the Convention and those of the Cartagena Protocol and Nagoya Protocol” so, we need to circumscribe to CBD, PCB, and NK objectives. Based on that, I firmly consider that the questions here are: how the technical information associated with SynBio that has been generated up to now is affecting in negative or positive ways the conservation of biological diversity? are there examples of the sustainable use of biodiversity of its components? Are there examples of fair and equitable sharing of benefits arising out the utilization of genetic resources? How do the mentioned SynBio products, that have been recently generated and mentioned here, adversely affect the conservation and the sustainable use of biodiversity? How the transboundary movement of the mentioned SynBio products could eventually be affecting other products?
I think we must take into account what has been already publish and differentiate it from what could eventually be possible to do with SynBio. The Dec. 14/19 and XII/24 do not pretend to solve all the issues associated with SynBio, just the ones related to CBD, PCB, and eventually NP.
For example, I have read here some comments about pharmaceuticals and this is explicitly excluded from CPB. The focus of CPB “apply to the transboundary movement, transit, handling and use of all living modified organisms that MAY HAVE ADVERSE EFFECTS on the conservation and sustainable use of biological diversity, taking also into account risks to human health”, so the question here could be how the SynBio products that are LMOs and have been reported in literature are having adverse effects on the conservation and sustainable use of biological diversity, and on human health?
Again thanks to all of you for your interesting comments.
Sincerely yours,
Pedro Rocha

1). New technological developments in synthetic biology since the last meeting of the Ad Hoc Technical Expert Group
Thank you Ms. Maria de Lourdes Torres for volunteering to moderate this discussion topic.  I’ll do my best to stay on topic.
By way of introduction, my name is Jim Louter from Canada working in the federal department of ‘Environment and Climate Change Canada’ where our small group has been conducting environmental risk assessments of living organisms since 1997 when our regulations were implemented.  This includes assessment of both naturally occurring organisms as well as those that have been genetically modified. ‘Synthetic biology’ is not a defining term in our regulatory scheme but rather, the application of science and engineering to the organism or to the use of the organism is. 

I believe the objective of this topic is to bring to mind any new developments that might change the way the discussion has been progressing since the last time experts met in 2017 in Montreal – I agree, as have many others, that while there has been continued evolution and refinement of methods, and no end of new proposals or ideas, there hasn’t really been any new development that would raise new issues not already considered.  As many Parties have already been asked to address a similar,  but more focussed on genome editing question, by written submission to the Secretariat, (“namely: New technological developments in synthetic biology since the last meeting of the Ad Hoc Technical Expert Group in December 2017, including the consideration, among other things, of concrete applications of genome editing if they relate to synthetic biology, in order to support a broad and regular horizon scanning process” as part of NOTIFICATION No. 2018-103 as part of a COP decision), I won’t repeat the submission made by Canada in this regard.

In replying to Claudia Vickers post #9383, I'm also supporting the publication of Australia of the Horizon Scanning document that ACOLA published last year.  I brought a copy of it with me to COP14 and to the Contact Group on Synthetic Biology as an example of what 'horizon scanning' on this topic would look like - it would make a lot of sense for us to build on the work done by others and not re-invent the wheel. 

thank you,

Jim Louter, Environment and Climate Change Canada.

Dear forum participants,
My name is Ana Atanassova, and I have been following the discussions under the CBD on risk assessment and synthetic biology over the last few years. My professional experience is in the area of biotech research from academia and ag biotech industry, as well as risk assessment of LMOs. I had the privilege to contribute to the work of the 2017 AHTEG on synthetic biology and am happy to address topic 1 – new technological developments in synthetic biology since the last meeting of the AHTEG.
As others have pointed out, the AHTEG report had listed a number of developments in modern biotechnology, and the current forum participants are adding numerous examples of new developments since the end of 2017.  These illustrate the inevitable progress in science and technology and cover varied examples ranging from hypothetical “new” applications, to early laboratory research, to re-branded “new” applications (e.g. genome editing for plant breeding), to concrete examples of exiting developments in medicine, agriculture, biosensing and fermentation. While the examples provided here, and in the information submissions (https://bch.cbd.int/synbio/submissions/) constitute an impressive list of references, it will be very helpful that these are examined in terms of the objectives of the CBD and its protocols, and as to whether the actual (and not hypothesized) developments challenge the existing provisions for risk assessment and risk management of the Cartagena Protocol. Similar point was eloquently made in post [#9456]. Therefore, while it is helpful and very informative to keep an eye and exchange information on scientific publications focusing on technological developments, it would be much more relevant to identify concrete and realistically foreseeable applications (products) and examine their potential positive and negative impacts vis-à-vis the three objectives of the Convention and those of the Cartagena Protocol and Nagoya Protocol.
Looking forward to the discussions for the rest of the month,
Best regards,
Ana

Dear all—

I am Bob Friedman, a researcher with the J. Craig Venter Institute (JCVI), a non-profit genomics research institute in the United States that has a very active synthetic biology program.  I was honored to serve on both the 2015 and 2017 Synthetic Biology AHTEGs and would like to offer my comments on new technological developments since the 2017 AHTEG meeting.

I, like many other participants on this Topic have observed, see few new technological developments since that time and believe that we are better served focusing on applications.  In this regard, the comments offered by Geoff Turner [#9449] on “technology readiness levels” were particularly helpful. 

For example, the eight base Hachimoji DNA referred to in several postings (citation in [#9382]), is most certainly a new technological development, but as of now, its status is that of a chemical that has never been incorporated into a living organism.  It is an impressive scientific achievement after years of effort, but it is a demonstration of a concept. 

Valeri Vasquez [#9407] comments on several new approaches for potentially limiting the geographic spread of gene drives.  Several groups are actively exploring these in the lab, with at least one laboratory demonstration of a successful “threshold” gene drive, i.e., one that requires a larger fraction of the population to contain the genetic alteration before it “drives”.  (Buchman, A., J. M. Marshall, D. Ostrovski, T. Yang and O. S. Akbari (2018). "Synthetically engineered Medea gene drive system in the worldwide crop pest Drosophila suzukii." Proc Natl Acad Sci U S A 115(18): 4725-4730.)  While additional work is needed to develop these from proof of concept to application, these illustrate the next level of technology readiness.

Applying this concept of technological readiness can help focus our attention to where it is needed most.

Regards,
Bob Friedman

This is Ryo Kohsaka from university in Japan.

I had the opportunity to participate in the past two AHTEGs.

Just one simple reminder that I suggested concrete technological elements which may fall under the "synthetic biology". The group as AHTEG agreed that  it would be less focusing on individual elements  given the speed of changes etc.

My apology for the delayed response due to my personal reasons.

Dear Margret
My name is Dan Tompkins, and I am one of a group of co-authors developing a technical assessment for the International Union for Conservation of Nature (IUCN) on “Genetic Frontiers for Conservation: An Assessment of Synthetic Biology and Biodiversity Conservation” (see https://www.iucn.org/theme/science-and-economics/our-work/other-work/synthetic-biology-and-biodiversity-conservation). I am just writing to let you know that the questions you raise with regards to wildlife and conservation have been considered in quite some depth in this assessment. While a draft version was previously online on the IUCN website (an ‘open review’ phase in which we received 700+ comments), the final version is planned to be available this month (with also all the review comments received, and how they were responded to for the final version).
Best wishes, Dan.

I have the same concern on the definition of 'synthetic biology'. Obviously the current one is a description other than a definition. I guess that is why the AHTEG was given the mandate to develop a suitable definition. From the view of a non-native English speaker, I assume that 'synthetic' points to non-nature made. Thus the synthetic target gene (e.g. xeno-) should be novel that is different from any existed one, which may be a way to distinguish it from LMOs.

Dear All
I would just like to support the point made in post #9462, that the eight base Hachimoji DNA concept work does count as a new technological development.
Regards, Dan Tompkins.

Dear Maria, Dear all,

Even though it has been stated several time in this forum that there has been no technological advancement since the last AHTEG, I would like to emphasize that we should not underestimate all the small technological advancements that are in sum remarkable.

I agree with previous contributions about the incredible dynamics in synthetic biology as summarized by Paul (#9409). The pace of technical developments in the field of biotechnology that are harnessed for synthetic biology is not slowing down. With third generation sequencing technologies from PacBio or Oxford Nanopore coming of age reading DNA is getting an additional boost. Importantly for synthetic biology also synthesis of DNA sequences is becoming cheaper and more efficient allowing longer fragments to be generated. Precise integration and manipulation of sequences is simplified by the ever-growing toolbox (including CRISPR), which is constantly applied to new target species for synthetic biology. Combined with the developments in computing, bioinformatics and data analysis the possibilities for automation and high throughput research in synbio are constantly increasing.

I think one of the major advancements is expected in the combination of those fast-developing fields. Progress should not be considered independently, but has to be evaluated viewing the overall picture.

All the best,
Margret

Dear all,
seconding both on Bobs (#9462) and Dans (#9466) response, we should see the eight base Hachimoji DNA as concept and not as new technological development at this stage.
And answering the main question of topic 1 I think that there are no new technological development but rather a rapid application of the existing as nicely illustrated by the many case studies listed above. Agreeing with Todd (#9395)- thus it is extremly important that we establish a effective horizon scanning process, continuously assessing the progress in the field.

Gernot

Dear Friends
I like to heartily expression of my gratitude and Thank you so much for the great efforts. I am continue following up these information  carefully and will come some of valuable contribution from IPLC perspective. 

Kamal Kumar Rai
IPLC
Nepal

Dear All - First I would like to congratulate and thank the Moderators for setting the questions for this discussion.
My name is Kelebohile Lekoape and I'm a Regulatory Manager for BASF in South Africa.  My biotech experience spans public reaserch, gvt, intergvtal orgs and industry.  I have had the opportunity to follow the CPB discussions for several years.
There seems to be a general consensus on there being no new technological developments since the last AHTEG 2 years ago [#9435, #9363, #9421, #9462] etc.  Everything that may be deemed new has relied on the same technologies and/or has not passed the proof of concept phase.  As a result, I'm inclined to agree with [#9449, #9460] that applications of synthetic biology that result in the production of LMOs ought to be excluded from this discussion because the CPB and national legislation would cover their regulation.  Mindful though that research is an evolving continuum, I would like to suggest that the discussion is rather focussed on the applications of current tools and whether or not derived products have an impact (positive or negative) on the objectives of the CBD.

Thank you All for the interesting inputs.

Kelebohile

Dear Forum Participants,

I would like to thank you so much for all your constructive and enlightened contributions to this forum. We have compiled a significant amount of technical information relevant to the topic of this forum.

The Secretariat will summarize all your contributions in a report that will be considered for the discussions of the AHTEG.

You have pointed out several key issues to be discussed: what the real new technological developments in synthetic biology in the last years are, which advances are proof of concepts, and which developments are applications of these technologies. The AHTEG should definitely consider these different approaches when analyzing this topic.

For us, it is very important to be aware of all the progress made, and that which is yet to come in the field of synthetic biology, keeping in mind the objectives of the Convention.

Again, many thanks for this fruitful conversation during this week.

Kind Regards

Maria

For info I include this reference and a commentary which are about SynBio.

Qian F. et al. (2019): Direct Writing of Tunable Living Inks for Bioprocess Intensification. Nano Letters | DOI: 10.1021/acs.nanolett.9b00066
Critical to the success of three-dimensional (3D) printing of living materials with high performance is the development of new ink materials and 3D geometries that favor long-term cell functionality. Here we report the use of freeze-dried live cells as the solid filler to enable a new living material system for direct ink writing of catalytically active microorganisms with tunable densities and various self-supporting porous 3D geometries. Baker's yeast was used as an exemplary live whole-cell biocatalyst, and the printed structures displayed high resolution, large scale, high catalytic activity and long-term viability. An unprecedented high cell loading was achieved, and cell inks showed unique thixotropic behavior. In the presence of glucose, printed bioscaffolds exhibited increased ethanol production compared to bulk counterparts due largely to improved mass transfer through engineered porous structures. The new living materials developed in this work could serve as a versatile platform for process intensification of an array of bioconversion processes utilizing diverse microbial biocatalysts for production of high-value products or bioremediation applications.

https://pubs.acs.org/doi/10.1021/acs.nanolett.9b00066

Anne M Stark, Lawrence Livermore National Laboratory: 3-D-printed live cells convert glucose to ethanol, carbon dioxide to enhance catalytic efficiency

https://phys.org/news/2019-03-d-printed-cells-glucose-ethanol-carbon.html#jCp

Dear forum participants,

I want to pick up and comment two major points from the discussions.

Discussion of the definition of synthetic biology is a reoccurring theme under the CBD since it sets the boundaries for discussions under the topics. This is complicated by mixing up concepts of synthetic biology (e.g. modular design and engineering of biological systems, xenobiology) with technologies drawn from genetic engineering, biotechnology, engineering, or chemistry, used to achieve particular functionalities (see Synthetic Biology Position Paper by German Research Foundation: bch.cbd.int/database/attachment/?id=18990). I believe that it is unhelpful to simply define synthetic biology in terms of a spectrum of methods. Given the difficulty of clearly defining the field, discussions often centre on a very broadly defined term that may encompass large areas or indeed all of genetic technology and molecular biology, leading to discussions and recommendations for synthetic biology that are redundant with respect to existing regulations. I suggest that discussions should rather focus on applications, which are based on synthetic biology concepts, but exclude as suggested by #9473 (also #9383) applications that result in the production of LMOs whose regulation is covered within the scope of the Cartagena Protocol and national legislation. At the current stage of applications, this appears to be more an issue of horizon scanning.

I agree with the many posts ([#9435, #9363, #9421, #9462, …] on this forum stating that since the 2017 AHTEG report there have been no conceptionally novel developments of technologies related to synthetic biology, but rather very dynamic advancements and broader applications of existing technologies, such as advancements in DNA synthesis and assembly, high throughput DNA engineering (DNA foundries), and non-natural nucleotides and amino acids (brought up e.g. by #9409, #9417, #9410).

Dear colleagues,

I also had the opportunity and the honor to participate in the past two AHTEGs and as a witness of a long and productive online discussion on the issue of Synthetic Biology definition as well as in the first AHTEG meeting which end at 10:30 pm the last day I support participants who ask not to reopen this discussion and to focus in the topics which are our mandate, although at the same time I see the merit of concerned colleagues.
Prof.Dr. O.A.EL-Kawy in post [#9448] is more specific “With regards to the definition of Synthetic biology, I believe that the SynBio AHTEG already proposed a definition and it was adopted by the COP Decision XII/17. Openning the discussion on Definition right now will just shift the forum away from it intended purpose,”

On the topic of new developments I would like to share the examples and the views expressed in post [#9416] by Dr. Eva Sirinathsinghji

Respectfully,

Dr Lazaro Regalado

Dear forum participants,
Thank you for your interventions. This topic is now closed for comments.

Secretariat

POSTED ON BEHALF OF Mr. Reynante Ordonio, Philippines (Note: this message arrived at the Secretariat before the closing of the discussion.)

I am Reynante Ordonio of the Philippine Rice Research Institute. It is an honor to take part in this interesting forum.
In the Philippines, we have recently made  a set of policy recommendations on how products of New Plant Breeding Techniques (NPBT) are going to be assessed, and part of the said techniques are Synthetic Biology and gene editing. We are pushing for case-by-case analysis of NBT products wherein those that ultimately will not have novel combination of materials will not be subject to the Cartagena Protocol (here, I call GM regulation) but to existing regulations on non-GMOs, and this is in agreement with what Andrew has pointed out (#9372).
In crafting the policy recommendations, we had to consider the following definition:
Synthetic genomics is “the engineering of biological components and systems that do not exist in nature and the re-engineering of existing biological elements; it is determined on the intentional design of artificial biological systems, rather than on the understanding of natural biology” (Synbiology, 2006). This involves the synthesis of complex DNA molecules that give rise to a functional gene or a complete genome, even without using any natural template (Lusser et al., 2011).

Based on this definition, I think that the Australian example mentioned in Louisa’s post (#9379) should not be considered Synthetic Biology since the genes were sourced out from naturally occurring genomes. The high oleic acid soybean developed by Calyxt and the canola/rapeseed by Cibus mentioned by Marie (#9369) are indeed not under SynBio since they are produced through NBT (TALENs and ODM, respectively).

It should be clear that DNA must be synthesized (without a natural template) to be considered synthetic (process-based classification). Based on a product-based approach of assessment, I believe that Synthetic Biology has the ability to produce products that can range from non-GMO (using principle of substantial equivalence; #9368), GMO, to partially and completely artificial organisms. I would love to discuss more about this particularly on the following cases:  (1) if a gene is synthesized to perfectly resemble (or almost) a naturally occurring gene;  (2) if a synthesized gene considerably differs from a naturally occurring gene; (3) if the synthesized gene is novel (in this case, not found in nature); and (4) if a novel genome is synthesized.