Topic 2: Experience with detecting and identifying newly developed and unauthorized LMOs
Dr. Raja Chalghoumi,
Tunisia#11946
Tunisia#11946
il y a 3 ans
Dear Colleagues,
Welcome to the online discussions of the Network of Laboratories for the Detection and Identification of Living Modified Organisms.
My name is Raja Chalghoumi, I am an agronomist with a Ph.D. in Agronomic Sciences and Bioengineering. Currently, I am an Associate Professor in Animal Sciences at the College of Agriculture - University of Carthage – Tunisia. Moreover, I am a certified professional in Biorisk Management and Biosecurity, a member of the Tunisian Biosafety Technical Commission, and a member of the Ad Hoc Technical Expert Group (AHTEG) on Risk Assessment and Risk Management
Under this second topic, I would like to gather information on your experiences detecting and identifying newly developed and unauthorized LMOs. From the discussions of the Network in 2019, detecting and identifying newly developed and unauthorized LMOs, as well as LMOs with unknown sequences, were noted as challenges for Parties. It was suggested that this could relate to new genetic elements and new species used to develop LMOs, among other things. Further, in decision CP-10/11, the COP-MOP recognized the need for capacity-building on detecting and identifying unauthorized LMOs. Given the recognized relevance of this topic, I would like to invite participants to share information related to the following:
- Could you describe your experience with the detection and identification of newly developed LMOs?
- Similarly, what experience can you share regarding the detection and identification of unauthorized LMOs?
- To overcome the challenges faced, what steps did you take, or analytical tools did you implement?
- What challenges do you foresee in the near future for these types of LMOs?
I look forward to your valuable interventions.
Raja Chalghoumi
Welcome to the online discussions of the Network of Laboratories for the Detection and Identification of Living Modified Organisms.
My name is Raja Chalghoumi, I am an agronomist with a Ph.D. in Agronomic Sciences and Bioengineering. Currently, I am an Associate Professor in Animal Sciences at the College of Agriculture - University of Carthage – Tunisia. Moreover, I am a certified professional in Biorisk Management and Biosecurity, a member of the Tunisian Biosafety Technical Commission, and a member of the Ad Hoc Technical Expert Group (AHTEG) on Risk Assessment and Risk Management
Under this second topic, I would like to gather information on your experiences detecting and identifying newly developed and unauthorized LMOs. From the discussions of the Network in 2019, detecting and identifying newly developed and unauthorized LMOs, as well as LMOs with unknown sequences, were noted as challenges for Parties. It was suggested that this could relate to new genetic elements and new species used to develop LMOs, among other things. Further, in decision CP-10/11, the COP-MOP recognized the need for capacity-building on detecting and identifying unauthorized LMOs. Given the recognized relevance of this topic, I would like to invite participants to share information related to the following:
- Could you describe your experience with the detection and identification of newly developed LMOs?
- Similarly, what experience can you share regarding the detection and identification of unauthorized LMOs?
- To overcome the challenges faced, what steps did you take, or analytical tools did you implement?
- What challenges do you foresee in the near future for these types of LMOs?
I look forward to your valuable interventions.
Raja Chalghoumi
1. Could you describe your experience with the detection and identification of newly developed LMOs?
- I have been detected 35S CaMV Promoter and NOS terminator in food and animal feed ingredients for 7 years ago.
2. Similarly, what experience can you share regarding the detection and identification of unauthorized LMOs?
- Never
3. To overcome the challenges faced, what steps did you take, or analytical tools did you implement?
- To use Real-time PCR and assign the Limit of Detection (LOD) at 0.01% or 0.1%
4. What challenges do you foresee in the near future for these types of LMOs?
- It is accepted by the people and can be consumed safely.
- I have been detected 35S CaMV Promoter and NOS terminator in food and animal feed ingredients for 7 years ago.
2. Similarly, what experience can you share regarding the detection and identification of unauthorized LMOs?
- Never
3. To overcome the challenges faced, what steps did you take, or analytical tools did you implement?
- To use Real-time PCR and assign the Limit of Detection (LOD) at 0.01% or 0.1%
4. What challenges do you foresee in the near future for these types of LMOs?
- It is accepted by the people and can be consumed safely.
Qsn 1 & 2: Experience with screening of LMOs
-Limited multiplex options available for screening at either of detection/identification/quantification levels where most available commercial kits would do P34S, P35S and TNOS (TaqMan detection option) or substitute one of them with ESPS. Any novel promoters or terminators outside these will be missed during screening due to the inability of the kit to reach and report on it.
-Costly to procure screening kits
-Protein-based LMO Screening kits are not as robust (LFD and ELISA). Usually the strips are sold as single combs with options of multiplexing being procuring separate events/targets for multi-combs.
-Sample prep difficulties for oily samples. Poor quality of nucleic acids from such extractions as in soy sauces.
Q3: Probable solutions
- Substituting kits with primer design for the events/targets to cut on cost (yet to try)
Q4: Prospective Challenges
-With the increase in events and lag in development of corresponding screening kits for the novelties, testing will not always be a reflection of the positivity of the sample if not done exhaustively for all the available market approvals.
-Limited multiplex options available for screening at either of detection/identification/quantification levels where most available commercial kits would do P34S, P35S and TNOS (TaqMan detection option) or substitute one of them with ESPS. Any novel promoters or terminators outside these will be missed during screening due to the inability of the kit to reach and report on it.
-Costly to procure screening kits
-Protein-based LMO Screening kits are not as robust (LFD and ELISA). Usually the strips are sold as single combs with options of multiplexing being procuring separate events/targets for multi-combs.
-Sample prep difficulties for oily samples. Poor quality of nucleic acids from such extractions as in soy sauces.
Q3: Probable solutions
- Substituting kits with primer design for the events/targets to cut on cost (yet to try)
Q4: Prospective Challenges
-With the increase in events and lag in development of corresponding screening kits for the novelties, testing will not always be a reflection of the positivity of the sample if not done exhaustively for all the available market approvals.
Dear Colleagues,
We believe that your expertise and contribution to the online forum would greatly enrich the discussions under the addressed topics. Your comments, insights, and tips would be very appreciated.
Best regards
We believe that your expertise and contribution to the online forum would greatly enrich the discussions under the addressed topics. Your comments, insights, and tips would be very appreciated.
Best regards
1. Could you describe your experience with the detection and identification of newly developed LMOs?
Answer: In our laboratory, we mostly go for the detection of P35S and tnos or other elements. But have no experience specific to the detection and identification of newly developed LMOs. This could be because we usually don’t receive any samples for analysis as import of LMOs (GMOs in viable form) are not allowed in Bhutan.
2. Similarly, what experience can you share regarding the detection and identification of unauthorized LMOs?
Answer: Since our agency (regulating the GMO/LMO) in Bhutan did not receive any applications, there are no approved GMOs as of now.
3. To overcome the challenges faced, what steps did you take, or analytical tools did you implement?
Not Applicable
4. What challenges do you foresee in the near future for these types of LMOs?
Answer: Generally, the challenges would be in quantification of the GMOs to see the compliance against the labelling requirements (as GMOs are allowed to enter the country in non-viable form for food and feed in the country). Also, it is difficult to fulfill the technical aspects of the ISO/IEC 17025 accreditation like the defining the LOD, LOQ of the machine, the measurement uncertainty, method validation, challenges in adopting new technologies in the area of GMO detection due to limited financial resources etc.
Answer: In our laboratory, we mostly go for the detection of P35S and tnos or other elements. But have no experience specific to the detection and identification of newly developed LMOs. This could be because we usually don’t receive any samples for analysis as import of LMOs (GMOs in viable form) are not allowed in Bhutan.
2. Similarly, what experience can you share regarding the detection and identification of unauthorized LMOs?
Answer: Since our agency (regulating the GMO/LMO) in Bhutan did not receive any applications, there are no approved GMOs as of now.
3. To overcome the challenges faced, what steps did you take, or analytical tools did you implement?
Not Applicable
4. What challenges do you foresee in the near future for these types of LMOs?
Answer: Generally, the challenges would be in quantification of the GMOs to see the compliance against the labelling requirements (as GMOs are allowed to enter the country in non-viable form for food and feed in the country). Also, it is difficult to fulfill the technical aspects of the ISO/IEC 17025 accreditation like the defining the LOD, LOQ of the machine, the measurement uncertainty, method validation, challenges in adopting new technologies in the area of GMO detection due to limited financial resources etc.
Dear colleagues,
My name is Daniela Wahler. I work for the German Federal Office of Consumer Protection and Food Safety (BVL) in the Unit traceability, detection methods, Biosafety Clearing-House. BVL is home to the competent national authority for the Cartagena Protocol and also to a working group that develops the official control methods for the detection and identification of LMO in Germany.
To my understanding newly developed LMO have genetic modifications that consist of rarely used genetic elements. Therefore, the modified sequences for many if not most newly developed LMO cannot be detected using standard screening approaches. The same problem is faced regarding numerous unauthorized LMO. Consequently, the increasing number of newly developed and unauthorized LMO has become a problem. To overcome this challenge several approaches are used in Germany:
1. The simplest and most obvious solution is to adapt the portfolio of standard screening methods. In Germany, new element- and/or construct-specific screening methods are developed and validated on a regular basis considering the prevalence of genetic elements and constructs in LMO. Respectively, the German table of screening methods is continuously being extended (https://euginius.eu/euginius/pages/methodSearch_searchview.jsf, select method set ABC).
2. In order to detect LMO that are not authorized in the EU but commercialized elsewhere, numerous officially validated methods have been developed.
3. As mentioned under topic 1 (post #11973), NGS approaches complement the toolbox for the detection and identification of newly developed and unauthorized LMO. Due to the high resolution of the technology they allow for the simultaneous detection of numerous known LMO, the further molecular characterization and identification of unauthorized LMO. Furthermore, the continually collected NGS data allows for the development of AI models to detect even unknown LMO.
In the near future, the biggest challenges will be related to the ever-increasing number and diversity of LMOs and, therefore, access to information and reference material to these LMO.
My name is Daniela Wahler. I work for the German Federal Office of Consumer Protection and Food Safety (BVL) in the Unit traceability, detection methods, Biosafety Clearing-House. BVL is home to the competent national authority for the Cartagena Protocol and also to a working group that develops the official control methods for the detection and identification of LMO in Germany.
To my understanding newly developed LMO have genetic modifications that consist of rarely used genetic elements. Therefore, the modified sequences for many if not most newly developed LMO cannot be detected using standard screening approaches. The same problem is faced regarding numerous unauthorized LMO. Consequently, the increasing number of newly developed and unauthorized LMO has become a problem. To overcome this challenge several approaches are used in Germany:
1. The simplest and most obvious solution is to adapt the portfolio of standard screening methods. In Germany, new element- and/or construct-specific screening methods are developed and validated on a regular basis considering the prevalence of genetic elements and constructs in LMO. Respectively, the German table of screening methods is continuously being extended (https://euginius.eu/euginius/pages/methodSearch_searchview.jsf, select method set ABC).
2. In order to detect LMO that are not authorized in the EU but commercialized elsewhere, numerous officially validated methods have been developed.
3. As mentioned under topic 1 (post #11973), NGS approaches complement the toolbox for the detection and identification of newly developed and unauthorized LMO. Due to the high resolution of the technology they allow for the simultaneous detection of numerous known LMO, the further molecular characterization and identification of unauthorized LMO. Furthermore, the continually collected NGS data allows for the development of AI models to detect even unknown LMO.
In the near future, the biggest challenges will be related to the ever-increasing number and diversity of LMOs and, therefore, access to information and reference material to these LMO.
Topic 2: Experience with detecting and identifying newly developed and unauthorized LMOs
- Could you describe your experience with the detection and identification of newly developed LMOs.
Answer: I am Dr. Bughe Rhoda Nsen, lecturer and manager of the Food and Drug Safety Laboratory, at the Biotechnology center, University of Yaounde 1, Cameroon. The Biotechnology Centre of the University of Yaounde I possesses the capacity to identify pathogenic microorganism in poultry, fish and shellfish, meat and water including LMOs using the molecular biology technologies.
- Similarly, what experience can you share regarding the detection and identification of unauthorized LMOs
Answer: As of now, research at the Biotechnology center of the University of Yaounde I has been strictly directed towards research carried out by Masters’ and Doctorate Students. These researches have not been focused on LMO detection.
- To overcome the challenges faced, what steps did you take, or analyticals tool did you implement.
Answer: not applicable
- What challenges do you foresee in the near future for these types of LMOs
Answer: In Cameroon, LMOs and derived product importation has been for field trials specifically on GM cotton in the Northern Regions of Cameroon. As of now, Cameroon has not received an official request for the importation of LMOs for food and feed. The lack of request for the importation of LMOs into the country coupled with the porous borders, constitutes a challenge in the control of LMOs introduction into the national territory. It is not surprising that, some LMOs or derived products may be found in the local market. Hence the need for testing/ detection in the markets and the entry ports.
- Could you describe your experience with the detection and identification of newly developed LMOs.
Answer: I am Dr. Bughe Rhoda Nsen, lecturer and manager of the Food and Drug Safety Laboratory, at the Biotechnology center, University of Yaounde 1, Cameroon. The Biotechnology Centre of the University of Yaounde I possesses the capacity to identify pathogenic microorganism in poultry, fish and shellfish, meat and water including LMOs using the molecular biology technologies.
- Similarly, what experience can you share regarding the detection and identification of unauthorized LMOs
Answer: As of now, research at the Biotechnology center of the University of Yaounde I has been strictly directed towards research carried out by Masters’ and Doctorate Students. These researches have not been focused on LMO detection.
- To overcome the challenges faced, what steps did you take, or analyticals tool did you implement.
Answer: not applicable
- What challenges do you foresee in the near future for these types of LMOs
Answer: In Cameroon, LMOs and derived product importation has been for field trials specifically on GM cotton in the Northern Regions of Cameroon. As of now, Cameroon has not received an official request for the importation of LMOs for food and feed. The lack of request for the importation of LMOs into the country coupled with the porous borders, constitutes a challenge in the control of LMOs introduction into the national territory. It is not surprising that, some LMOs or derived products may be found in the local market. Hence the need for testing/ detection in the markets and the entry ports.
Dear Colleagues,
In our laboratory (Ministry of Agriculture, Brazil), we perform screening methods based on seven different GMO sequences in order to identify possible events in a sample, in case of positive results we confirm the LMO by event specific methods analysis. Considering unauthorized LMOs, the strategy of analysis is established in a case-by-case scenario, depending on the biological species and the origin of the material. Newly developed LMO are authorized according to the evaluation by the National Technical Commission for Biosecurity (CTNBio) and the applicants should provide methodology and reference materials to the Official Laboratory aiming at method validation and market control.
Best regards,
Cairo de Oliveira
In our laboratory (Ministry of Agriculture, Brazil), we perform screening methods based on seven different GMO sequences in order to identify possible events in a sample, in case of positive results we confirm the LMO by event specific methods analysis. Considering unauthorized LMOs, the strategy of analysis is established in a case-by-case scenario, depending on the biological species and the origin of the material. Newly developed LMO are authorized according to the evaluation by the National Technical Commission for Biosecurity (CTNBio) and the applicants should provide methodology and reference materials to the Official Laboratory aiming at method validation and market control.
Best regards,
Cairo de Oliveira
Our current detection method is based on 35S promoter and NOs terminator. We do not have any experience in detection and identification of unauthorised LMOs. The key challenges in the near future for these types of LMOs in the face of detection capacity gaps is that they may lead to false negatives that may result in loss of trade opportunities and even loss of biodiversity. This is much more critical for most African countries that have porous borders with free movement of agricultural produce between countries through unchartered routes
Dear Mtnonga,
Thank you for sharing. Most African countries LMO detection method is 35S promoter and NOs terminator based.
Please see this published paper for more experience which is done in Ethiopia Environment Protection authority, GMO Detection and Research Lab.
https://www.opastpublishers.com/peer-review/detection-for-suspected-genetically-modified-maize-and-soybean-crops-in-the-selected-places-of-ethiopia-5325.html
Thank you for sharing. Most African countries LMO detection method is 35S promoter and NOs terminator based.
Please see this published paper for more experience which is done in Ethiopia Environment Protection authority, GMO Detection and Research Lab.
https://www.opastpublishers.com/peer-review/detection-for-suspected-genetically-modified-maize-and-soybean-crops-in-the-selected-places-of-ethiopia-5325.html
Dear Mr. Mtnonga, Mr. Adugnaw, and Mr. de Oliveira,
Thank you very much for sharing with us information about the situations in your respective countries.
Best regards
Thank you very much for sharing with us information about the situations in your respective countries.
Best regards
Dear Colleagues,
I would like to gently remind you that the forum will close on Tuesday, 28 November 2023 at 17:00 EST (UTC -5).
We look forward to your contributions and exchanges!
Best regards
I would like to gently remind you that the forum will close on Tuesday, 28 November 2023 at 17:00 EST (UTC -5).
We look forward to your contributions and exchanges!
Best regards
Dear Colleagues,
In México at CENAM the National Metrology Institute we have developed and certified reference materials to measure p35S and tNOS. https://www.cenam.mx/materiales/materiales.aspx?Descripcion=&Clave=&uso=0&division=600&B1=Buscar.
We have presented the following publications:
Copy number ratios determined by two digital polymerase chain reaction systems in genetically modified grains. Metrologia 51 (2014) 61–66 doi:10.1088/0026-1394/51/1/61.
Development, Optimization, and Evaluation of a Duplex Droplet Digital PCR Assay To Quantify the T‑nos/hmg Copy Number Ratio in Genetically Modified Maize. DOI: 10.1021/acs.analchem.5b03238
Anal. Chem. 2016, 88, 812−819.
Zea maize reference materials for genetically modified organism detection in Mexico. Ecology and evolution. 2019. DOI: 10.1002/ece3.5667
with best regards
Melina Pérez Urquiza
In México at CENAM the National Metrology Institute we have developed and certified reference materials to measure p35S and tNOS. https://www.cenam.mx/materiales/materiales.aspx?Descripcion=&Clave=&uso=0&division=600&B1=Buscar.
We have presented the following publications:
Copy number ratios determined by two digital polymerase chain reaction systems in genetically modified grains. Metrologia 51 (2014) 61–66 doi:10.1088/0026-1394/51/1/61.
Development, Optimization, and Evaluation of a Duplex Droplet Digital PCR Assay To Quantify the T‑nos/hmg Copy Number Ratio in Genetically Modified Maize. DOI: 10.1021/acs.analchem.5b03238
Anal. Chem. 2016, 88, 812−819.
Zea maize reference materials for genetically modified organism detection in Mexico. Ecology and evolution. 2019. DOI: 10.1002/ece3.5667
with best regards
Melina Pérez Urquiza
Dear colleagues,
My name is Emmanuel González-Ortega. I’m trained in Biotechnology (Engineering, Ms Sc, PhD), and I have expertise in LMO detection and identification, biosafety governance, molecular biology, risk assessment, socioeconomic considerations of LMO, synthetic biology. Our research group have investigated and published on the presence of unauthorized genetically modified maize in several Mexican regions, and on the presence of genetically modified maize in highly consumed food manufactured with maize in Mexico. I have previously participated in several fora in the Cartagena Protocol of the CBD. I would like to thank the Secretariat of the CBD for the organization of the on-line forum of the Network of Laboratories for the Detection and Identification of Living Modified Organisms and for the opportunity to participate in this discussion.
Regarding the second topic proposed for this on-line forum post [#11946], here are my considerations.
Based on previous posts by participants of the online forum [#11951] [#11959] [#11964] the reality of the state of LMO detection techniques and protocols in different countries has been made explicit, and consequently, the need to update LMO detection and capacity-building methodologies considering new generation LMOs but without ceasing to monitor LMOs from previous generations is evident.
I completely agree with Ms Wahler [#11974] regarding the impossibility of detecting new LMOs with previous and consolidated approaches and protocols for the detection and identification of LMOs (conventional PCR, Lateral Flow devices, ELISA tests), which is a problem for the biosafety governance of new and unauthorized LMOs. However, I think that the problem is more complex than the mere need to update the techniques used for monitoring and detection new LMOs. There are transversal issues implying politics, regulations, etc., for example, the heated debate in regions (i.e the European Union) on the exemption from the regulations, labeling and transparency to farmers and consumers of the presence of new LMO generated by genome editing (i.e. CRISPR-Cas) and their products, although in advance, genetically modified organisms already exist on the market through genome editing.
The intra- and inter-government regulation of the new generation LMOs (within the framework of the Cartagena protocol and the CBD) should be a fundamental initiative for strengthening the channels for the exchange of technical, technological and methodological information between countries that strengthen the biosafety governance and next-generation LMO detection.
Assuming that most countries do not have built-capacities for the detection of new generation LMOs, I consider that aspects and actions focused on achieving the consolidation of detection methods for new LMOs should be further discussed. Additionally, it is essential to establish and strengthen frames and channels of transparency on the information that biotechnology companies, research centers and universities provide to regulatory bodies and governments regarding: the release of information without restrictions on the genomes of modified organisms, the unrestricted release of the information on technologies used to modify the LMO, the release of unrestricted information on the methodologies used in the analysis, verification and validation of the modification produced in the LMO. The availability of this information, and additional topics that other experts attending this online forum would consider will be a significant step towards updating and consolidating the governance of biosafety of emerging LMO.
There is an outstanding meta-analysis carried out by Chu and Agapito-Tenfen on the unintended genomic effects of current and emerging genetic modification techniques (CRISPR-Cas) [DOI: https:// doi.org/10.3390/plants11212997 ]. In this study, the authors reviewed the analytical methods used to detect the unconsidered outcomes of genetic modification using CRISPR-Cas. It was found that the majority (approximately 26%) of the studies analyzed in the study found that PCR-based methods were used to identify the modifications made by CRISPR. Secondly, some sequencing technique variant was used (approximately 17%) to detect CRISPR-produced genetic modifications. “Indirect” techniques for detecting genetic modifications (i.e. mass spectroscopy, electrophoresis, microscopy, enzymatic assays, statistical analysis) were also used in a very marginal way (cumulative average approximately 8.2%). Although methods for detecting genetic modifications (intentional and unintentional) are mostly identified through sequencing methods, other techniques are used that could disperse information about the creation of LMOs through new genetic modification techniques and, consequently, about the potential impacts on biodiversity and human health.
Best regards,
Emmanuel.
My name is Emmanuel González-Ortega. I’m trained in Biotechnology (Engineering, Ms Sc, PhD), and I have expertise in LMO detection and identification, biosafety governance, molecular biology, risk assessment, socioeconomic considerations of LMO, synthetic biology. Our research group have investigated and published on the presence of unauthorized genetically modified maize in several Mexican regions, and on the presence of genetically modified maize in highly consumed food manufactured with maize in Mexico. I have previously participated in several fora in the Cartagena Protocol of the CBD. I would like to thank the Secretariat of the CBD for the organization of the on-line forum of the Network of Laboratories for the Detection and Identification of Living Modified Organisms and for the opportunity to participate in this discussion.
Regarding the second topic proposed for this on-line forum post [#11946], here are my considerations.
Based on previous posts by participants of the online forum [#11951] [#11959] [#11964] the reality of the state of LMO detection techniques and protocols in different countries has been made explicit, and consequently, the need to update LMO detection and capacity-building methodologies considering new generation LMOs but without ceasing to monitor LMOs from previous generations is evident.
I completely agree with Ms Wahler [#11974] regarding the impossibility of detecting new LMOs with previous and consolidated approaches and protocols for the detection and identification of LMOs (conventional PCR, Lateral Flow devices, ELISA tests), which is a problem for the biosafety governance of new and unauthorized LMOs. However, I think that the problem is more complex than the mere need to update the techniques used for monitoring and detection new LMOs. There are transversal issues implying politics, regulations, etc., for example, the heated debate in regions (i.e the European Union) on the exemption from the regulations, labeling and transparency to farmers and consumers of the presence of new LMO generated by genome editing (i.e. CRISPR-Cas) and their products, although in advance, genetically modified organisms already exist on the market through genome editing.
The intra- and inter-government regulation of the new generation LMOs (within the framework of the Cartagena protocol and the CBD) should be a fundamental initiative for strengthening the channels for the exchange of technical, technological and methodological information between countries that strengthen the biosafety governance and next-generation LMO detection.
Assuming that most countries do not have built-capacities for the detection of new generation LMOs, I consider that aspects and actions focused on achieving the consolidation of detection methods for new LMOs should be further discussed. Additionally, it is essential to establish and strengthen frames and channels of transparency on the information that biotechnology companies, research centers and universities provide to regulatory bodies and governments regarding: the release of information without restrictions on the genomes of modified organisms, the unrestricted release of the information on technologies used to modify the LMO, the release of unrestricted information on the methodologies used in the analysis, verification and validation of the modification produced in the LMO. The availability of this information, and additional topics that other experts attending this online forum would consider will be a significant step towards updating and consolidating the governance of biosafety of emerging LMO.
There is an outstanding meta-analysis carried out by Chu and Agapito-Tenfen on the unintended genomic effects of current and emerging genetic modification techniques (CRISPR-Cas) [DOI: https:// doi.org/10.3390/plants11212997 ]. In this study, the authors reviewed the analytical methods used to detect the unconsidered outcomes of genetic modification using CRISPR-Cas. It was found that the majority (approximately 26%) of the studies analyzed in the study found that PCR-based methods were used to identify the modifications made by CRISPR. Secondly, some sequencing technique variant was used (approximately 17%) to detect CRISPR-produced genetic modifications. “Indirect” techniques for detecting genetic modifications (i.e. mass spectroscopy, electrophoresis, microscopy, enzymatic assays, statistical analysis) were also used in a very marginal way (cumulative average approximately 8.2%). Although methods for detecting genetic modifications (intentional and unintentional) are mostly identified through sequencing methods, other techniques are used that could disperse information about the creation of LMOs through new genetic modification techniques and, consequently, about the potential impacts on biodiversity and human health.
Best regards,
Emmanuel.
Dear colleagues,
I am Litao Yang, a professor from Shanghai Jiao Tong UNiversity. In my group, we are working on developing new techniques for identifiation and quantification of LMOs.
In 2013, our group developed Tran-Seq based on NGS technique, which contains three modules for GMO analysis (Scientific Rep. 2013. 3: 2839). Module 1 was designed for use when the DNA sequence of the transformation vector is available (ISK-class 2). Module 2 was designed for use when the DNA sequence database of genetic elements and transgenic constructs from known GMOs are available and can be used as reference libraries. Module 3 was designed for use when a priori knowledge of the DNA sequence of the transformation vector or transgenic constructs is not available. For application of any of the three modules the non-GM genome sequence must be available as a reference. In 2022, we developed LIFE-seq, which combined capture enrichment and PacBIo sequencing for revealing the whole integration of exogenous DNA insertion. In LIFE-seq, one universal tilling probe library was constructed, which contained often used universal elements, mark genes, exogenous genes, and GM events. This library can be used for unknown GMO contents analysis. (plant biotechnology journal, 2022, 20(5):964-976 )
I am Litao Yang, a professor from Shanghai Jiao Tong UNiversity. In my group, we are working on developing new techniques for identifiation and quantification of LMOs.
In 2013, our group developed Tran-Seq based on NGS technique, which contains three modules for GMO analysis (Scientific Rep. 2013. 3: 2839). Module 1 was designed for use when the DNA sequence of the transformation vector is available (ISK-class 2). Module 2 was designed for use when the DNA sequence database of genetic elements and transgenic constructs from known GMOs are available and can be used as reference libraries. Module 3 was designed for use when a priori knowledge of the DNA sequence of the transformation vector or transgenic constructs is not available. For application of any of the three modules the non-GM genome sequence must be available as a reference. In 2022, we developed LIFE-seq, which combined capture enrichment and PacBIo sequencing for revealing the whole integration of exogenous DNA insertion. In LIFE-seq, one universal tilling probe library was constructed, which contained often used universal elements, mark genes, exogenous genes, and GM events. This library can be used for unknown GMO contents analysis. (plant biotechnology journal, 2022, 20(5):964-976 )
Could you describe your experience with the detection and identification of newly developed LMOs?
The CNRIBA has more than 10 years of experience in the analysis of LMO detection, however in recent years we have observed variations in the amplification cycles of endogenous genes.
- Similarly, what experience can you share regarding the detection and identification of unauthorized LMOs?
It is extremely complicated to detect unauthorized LMOs because the construction of the event is not known, in that sense this laboratory has been limited.
- To overcome the challenges faced, what steps did you take, or analytical tools did you implement?
We are currently looking for training to learn about new biotechnological techniques.
- What challenges do you foresee in the near future for these types of LMOs?
The methodologies that we currently use for the detection of LMOs will not be sufficient, one answer is sequencing, however the cost of its implementation will be high.
The CNRIBA has more than 10 years of experience in the analysis of LMO detection, however in recent years we have observed variations in the amplification cycles of endogenous genes.
- Similarly, what experience can you share regarding the detection and identification of unauthorized LMOs?
It is extremely complicated to detect unauthorized LMOs because the construction of the event is not known, in that sense this laboratory has been limited.
- To overcome the challenges faced, what steps did you take, or analytical tools did you implement?
We are currently looking for training to learn about new biotechnological techniques.
- What challenges do you foresee in the near future for these types of LMOs?
The methodologies that we currently use for the detection of LMOs will not be sufficient, one answer is sequencing, however the cost of its implementation will be high.
Dear participants and distinguished colleagues,
My name is Nancy Serrano-Silva, and I am affiliated with the Executive Secretariat of the Interministerial Commission for the Biosafety of Genetically Modified Organisms in Mexico. I am pleased to join this significant forum on the experience in the detection and identification of recently developed and unauthorized LMOs, within the context of the meeting of the Parties to the Cartagena Protocol on Biosafety.
I fully agree with the evident need to update detection methodologies and build capacities for the next generation of LMOs, while not neglecting the monitoring of those from previous generations. However, as highlighted before, the challenge extends beyond mere technical updates. We are confronted with overarching issues involving policies and regulations, such as the labeling of new LMOs generated by CRISPR-Cas. The presence of these organisms in the market before clear regulation underscores the urgency of addressing these issues on a global scale. In this regard, the intra- and intergovernmental regulation of next-generation LMOs, within the framework of the Cartagena Protocol and the Convention on Biological Diversity, must become a central initiative. Strengthening channels for the exchange of technical, technological, and methodological information among countries is essential for enhancing biosafety governance and the detection of next-generation LMOs.
I appreciate the opportunity to participate in this forum and look forward to contributing to an enriching dialogue on these crucial matters.
Sincerely,
Nancy
My name is Nancy Serrano-Silva, and I am affiliated with the Executive Secretariat of the Interministerial Commission for the Biosafety of Genetically Modified Organisms in Mexico. I am pleased to join this significant forum on the experience in the detection and identification of recently developed and unauthorized LMOs, within the context of the meeting of the Parties to the Cartagena Protocol on Biosafety.
I fully agree with the evident need to update detection methodologies and build capacities for the next generation of LMOs, while not neglecting the monitoring of those from previous generations. However, as highlighted before, the challenge extends beyond mere technical updates. We are confronted with overarching issues involving policies and regulations, such as the labeling of new LMOs generated by CRISPR-Cas. The presence of these organisms in the market before clear regulation underscores the urgency of addressing these issues on a global scale. In this regard, the intra- and intergovernmental regulation of next-generation LMOs, within the framework of the Cartagena Protocol and the Convention on Biological Diversity, must become a central initiative. Strengthening channels for the exchange of technical, technological, and methodological information among countries is essential for enhancing biosafety governance and the detection of next-generation LMOs.
I appreciate the opportunity to participate in this forum and look forward to contributing to an enriching dialogue on these crucial matters.
Sincerely,
Nancy
Dear colleagues from Mexico (Dr. Serrano Silva, Dr. Barrera Andrade and Dr. Gonzalez-Ortega),
Thank you for sharing information on the situation in your country and for your valuable exchange.
All the best
Thank you for sharing information on the situation in your country and for your valuable exchange.
All the best
