Team:Leiden/Human Practices
<!DOCTYPE html>
Integrated Human Practices
As part of our human practices, our team sought to view our project from a variety of perspectives, including societal and scientific needs, business point of view and what is legally required for the use of genetically modified organisms (Figure 1). On this page, we will report on the insights gained from interactions with regulatory, industrial and academic stakeholders. We have shown how our initial project concept was shaped by the stakeholders' expertise and knowledge.
-
Responsibility gap
We define genetically modified organisms (GMOs) as organisms, with the exception of human beings, in which the genetic material has been altered in a way that does not occur naturally by mating and/or natural recombination. They can have great potential, but it is still not allowed to use them outside of the laboratory. Therefore, we aimed to safeguard the release of GMOs. As we dug deeper into the literature and spoke with experts, we realized that there was a need for a standardized biocontainment system as well as a bridge between academia, policymakers and the industry for biosafety.
Input
Britte Bouchaut, Ph.D. Candidate Safe-by-Design for inherently safe Biotechnologies at TU Delft
Ms. Bouchaut told us that it would be difficult to perform a risk analysis on a large scale for the introduction of a GMO in the environment because strict legislation in Europe allows very little. To bridge this gap, policies must be more flexible and researchers should get more involved in the safety aspects of the process. Outside of academia, there is little interest in validating biocontainment systems, owing to a lack of immediate benefits and a lack of implementable applications.
Amalia Kallergi, Postdoctoral Researcher in the Biotechnology & Society group, Department of Biotechnology, TU Delft
Dr. Kallergi also mentioned that the validation of a biocontainment system is a major barrier in the progress of biosynthetic implementation. She explained that little research has been done on biocontainment systems outside of the lab partly due to relatively strict regulation, but mostly due to the absence of a responsible party to test a biocontainment system. Ironically, this lack of information is also one of the reasons why the regulation is strict: it is a vicious circle caused by the so-called responsibility gap. Simply said, there is no responsible actor in the transition from a proof of concept to a mature technology.
Enrique Asin Garcia, Ph.D. candidate in the NWO SafeChassis project. Synthetic biologist and biotechnologist studying biosafety mechanisms and related concepts at Wageningen University of Research
The responsibility gap was also mentioned by Enrique Asin Garcia. He also told us that, currently, the risks of engineered bacteria and biocontainment strategies are not assessed outside of the lab in field tests and potential application settings. There is a lack of metrics and standardized procedures for this and also a lack of knowledge regarding data collection since the strain engineers are not completely sure of how to measure safety and which levels of biosafety will be required for an application. To solve this issue, there is a huge need for tools for a standardized assessment, which is more a regulatory and political issue.
Professor Tom Ellis, Professor in Synthetic Genome Engineering at Imperial College London
Prof. Ellis mentioned that a single uncomplicated biocontainment method is probably the best option to be approved for use by regulators. He explained that complex biocontainment systems complexity can be detrimental to cell viability due to the increased need for resources for the cell, for example. Having plasmids in high copy numbers will cost a lot of resources to maintain both plasmids. For example, expressing multiple proteins (like a toxin antitoxin system) in high concentrations inevitably places a metabolic burden on the cell. This can potentially make it a challenge to express large gene clusters in high concentrations.
Tessa Alexanian, Safety and Security Program Officer at the iGEM Foundation
Ms. Alexanian confirmed to us that safety is of high importance in the iGEM community. Current biocontainment systems are not sufficient, because people are not using them. She hypothesizes that it is too complex for other iGEM teams to include a biocontainment system in the process from the beginning. Therefore, she thinks that there is a need for a biocontainment system that is simple to implement into an iGEM project, for example with interchangeable biobricks.
Integration
GMOs can be used for biomedical, industrial and environmental applications [1]. In addition, GMOs have become more common in many science and engineering sectors as powerful research tools for understanding and engineering biological processes and principles. Although some GMOs might be safe for release due to their unfavorable cellular fitness and engineering, biocontainment methods are still required to limit the proliferation of GMOs [1]. Especially when GMOs evidently outcompete naturally occurring species and harm the environment and/or human health. Due to these potentially disastrous risks , current regulations are very strict. Only when it is proven that there are no risks for the use of GMOs in the environment is it regarded as safe, as described in the precautionary principle [2].
Three main stakeholders can be identified in the development and implementation of synthetic biology applications. These stakeholders are scientists, industry and regulators, which have their own roles in the responsibility gap (Figure 2). Ms. Kallergi and Mr. Asin Garcia told us more about the identified responsibility gap. We realized that it is also important to bridge the responsibility gap. Therefore we aimed to extend the project beyond just synthetic biology. In our entrepreneurship plan, we consider the required social and regulatory matters to eventually bridge this gap.
Figure 2: Representation of the three stakeholders in the responsibility gap.
Ms. Alexanian and Prof. Ellis told us that if we wanted to develop a biocontainment system, it had to be a simple system to implement. In literature, we additionally found that most biocontainment systems have flaws that preclude them from being extremely effective protections [1]. This prompted us to develop a simple mechanism that could improve biosafety even further.
In the design of our system, we adapted the advice from Ms. Alexanian and Prof. Ellis to develop a simple easy-to-apply biocontainment system. Our goal is to provide a safe-by-design, plasmid-based biocontainment system , which can be implemented for various biotechnological applications. Therefore, we decided to make our final system called DOPL LOCK compatible with the standardized Standard European Vector Architecture (SEVA) plasmid nomenclature [3]. The SEVA plasmids are an already established standardized way of plasmid design, which will make it easier to implement biosecurity in all applications. Additionally, to offer a solution for a broad range of applications, we designed our toxin antitoxin (TA) system with inducible promoters. This way, we can redesign our system in such a way that it fits the needs of a specific application.
-
Science
We discovered that a biocontainment system was required, but it had to be designed as a simple system. Next, we wanted to further fine-tune our experimental design. Therefore we talked to various scientists to make decisions on our experiments.
Input
Renée Kapteijn, Ph.D. candidate at Leiden University
Ms. Kapteijn said that conjugation and transformation could take place with the same type of other compatible bacteria, but measuring this would be time-consuming in our limited lab time since horizontal gene transfer (HGT) takes place only under optimal circumstances. She also mentioned that an inducible promoter can be used for the proof-of-concept, proving that the cell is transformed with the plasmid and that the toxin works. The bigger the plasmid makes it more difficult to transform, thus keeping it to the essentials.
Professor Tom Ellis, Professor in Synthetic Genome Engineering at Imperial College London
In the design of our experiments, we wanted to integrate one plasmid into the genome. Prof. Ellis informed us that integrating one plasmid in the genome would not work because you then need the other plasmid in the cell the entire time. Ellis mentioned that this is unlikely cell engineering.
Dr. Erik Vijgenboom, Assistant Professor at Leiden University
With dr. Vijgenboom we discussed how we could control the toxin expression. He suggested using an inducible promoter. He also mentioned the importance of choosing the plasmids to co-transform, as the backbones and copy numbers of each plasmid are different. He also said that possible aspects of biocontainment systems that can be improved are preventing horizontal gene transfer in DNA in the biocontainment system.
Nathan Fraikin, Ph.D. candidate at Université Libre de Bruxelles
From Mr. Fraikin, we learned that optical density obtained by the plate reader in our experiments was not the best measurement to assess the toxicity of toxins, because bacteria may form resistance rapidly and the population could be outcompeted by resistant bacteria after long term incubation in LB media. Therefore, it was advised to evaluate toxicity based on colony forming units. Additionally, he mentioned that nuclease toxins were better toxin candidates for the DOPL LOCK system, as they would also digest DNA released after cell lysis and prevent HGT further. Lastly, he noted Ribosome binding site (RBS) sequences are another important factor of toxin and antitoxin gene expression, which are worth further investigation.
Akos Nyerges, Ph.D., Research Fellow at Harvard Medical School
With Akos, we talked about how our DOPL LOCK system could be potentially bypassed in living cells. He mentioned that the plasmids could merge and that bacteria might have ways to bypass the effects of the toxins by transposable elements [4] or point mutations. Also, he emphasized that there are currently no built-in ways to select for expression of the toxin gene in our system. Additionally, he said that we could use mutator strains to accelerate the evolution of our toxin modules and assess ways of escape. Mutator Escherichia coli (E. coli) strains have dramatically higher mutation rates than wild-type cells and therefore evolve faster [5]. In this way, we can test the durability of the DOPL LOCK system under simulated long-term evolution.
Integration
With Dr. Vijgenboom, Ms. Kapteijn and Prof. Ellis, we discussed the experimental details and used their feedback in our experimental design.
First, Dr. Vijgenboom emphasized the use of optimized plasmid backbones to get higher efficiency of gene expression in co-transformation. Following his suggestion, we dived into literature and found that the origin of replications (Oris) and the antibiotics selection markers are both crucial components in the plasmid backbone that influence its gene expression [6]. Oris strongly determine the compatibility of plasmids (two coresident plasmids to be stably inherited in the absence of external selection), especially when they are co-transformed [7]. We integrated this in our design by choosing the JUMP plasmid collection [6], which contained four kinds of Oris and two kinds of antibiotics selection markers, to perform co-transformation assays. From the results, we spotted the most compatible pair of plasmid backbones to optimize the plasmid gene expression as suggested.
Second, Ms. Kapteijn and dr. Vijgenboom told us that an inducible promoter could be used for the proof-of-concept, proving successful transformation with the plasmid and proving that the toxin works. By using an inducible promoter, we could further control the toxin. We incorporated this into our research lines and used the inducible promoter in the proof-of-concept.
Third, an early plan was to integrate one of our plasmids into the genome of E. coli strain BW23474 [8]. This turned out to be unfeasible in our limited lab time. With the feedback we got from Prof. Ellis, we quit this research line and focused more on our other research lines.
Lastly, with Ms. Kapteijn, we also discussed the possibility of horizontal gene transfer (HGT) experiments. At first, we wanted to show that HGT does not take place in our DOPL LOCK system. But, the difficulty of proving that HGT does not occur, did lead us to end the experiments involving this and focus on another part of the project. She told us that we could claim that the chance that both plasmids transfer via HGT to other cells is almost zero, since the frequency with which HGT takes place with two plasmids is low.
We also spoke to two researchers about the future development of our project. With Mr. Nyerges, we discussed potential design flaws of the current DOPL LOCK system and future improvements of our current system. In our design, there is no selection for toxin expression. This could potentially be solved by the elimination of an essential gene from the host. Subsequently, this essential gene would be co-expressed with the toxin under the control of the same promoter. Additionally, we talked about how transposable elements can integrate into the coding sequence of the toxin and therefore eliminate the toxic effect. We found that this problem can be overcome by using the E. coli strain W3110, which lacks transposable genetic elements [4]. If you want to read more on this topic, we would like to refer you to the Safety page. We also took the feedback from Mr. Fraikin into consideration for the future development of our project. He suggested applying colony forming units instead of optical density for future toxicity assays. He also advised us to put more effort into investigating the origins , the action of mechanisms and the gene regulations of TA systems if we want to broaden the application of our system. In this way, we may find more appropriate TA systems for future implementation. For example, nuclease-based toxins are the most suitable toxins for preventing HGT. For more information on this, we would like to refer to the Result page.
-
Application
The initial stages of our project indicated that there is a need for a simple biocontainment system. A biocontainment system could be used for different applications. In the search for a suitable application, we reached out to different companies and biotechnology experts. We wanted to discuss possible applications of our system and the considerations we needed to take into account when designing our system.
Input
Professor Jack Pronk, Full Professor and Head of Biotechnology Department, TU Delft
Our initial project idea was to use our system to prevent the spread of antibiotic resistance genes. When we talked to professor Pronk, he strongly discouraged us from choosing this as an application. He said that antibiotic resistance genes are not used to construct GMOs for industrial application. Besides, if there are antibiotic resistance genes used, these would not confer resistance to compounds used for the treatment of humans or animals. There are currently much better options than using antibiotic resistance genes, such as using CRISPR-Cas9 for genomic integration.
Robert Wagenveld, CEO at QM Environmental International B.V.
From our talks with Prof. Pronk, we realized that our system would mainly be applied to environmental biotechnology. Mr. Wagenveld, CEO of a bioremediation company, said that GMOs are not used in the field of environmental biotechnology because companies are not allowed to do so. In addition, he mentioned that they use many bacterial strains that break down various compounds. However, he mentioned that polyfluoroalkyl substances (PFAS) are detrimental for wastewater treatment and bioremediation. Robert said that currently, physical and chemical separation methods are used for extracting PFAS. However, these techniques are not sufficient enough as they do not break down PFAS but only separate it from other components. In the end, the amount of PFAS keeps piling up. Yi et al. (2016) have discovered a naturally occurring bacterium that can break down PFAS in certain circumstances [9]. A major drawback is that the culture conditions are very difficult to simulate in practice. Mr. Wagenveld said that GMOs could be the solution to the PFAS problem since you can then engineer a bacterium that breaks down PFAS in an efficient way and also grows in less extreme conditions.
Braden Gilbert, Vice-president of Micro-Bac International
We also talked to another bioremediation company: Micro-Bac. Gilbert said that they are not working with GMOs for three reasons. The first one is regulation: It is not allowed to introduce GMOs in the environment without a license. The second one is a lack of education: people are afraid of using GMOs. The last one is economics: it can be expensive to develop GMOs, among other things by applying for a license which takes a lot of time. He also mentioned that in the future, the number of substances that cannot be degraded by naturally occurring microbes will increase. We concluded that GMOs are necessary to effectively remove these pollutants.
Professor Tom Ellis, Professor in Synthetic Genome Engineering at Imperial College London
We discussed what criteria make a sufficient biocontainment system with Prof. Ellis. Tom mentioned that a good biocontainment system should be (i) simplicity of action, so no complex genetic circuits are involved in the containment strategies, (ii) easy to select for and (iii) not prone to mutations. This would mean that no special medium requirements are needed for the application of our biocontainment strategy. In addition, he mentioned three areas that need a proper biocontainment system: (i) gut and skin microbiome applications, (ii) water and environment remediation applications and (iii) algal biofuel-producing cells grown in open water ponds. He also said that our system is likely to provide enough safety to release GMOs in (semi-)contained environments.
Dr. Ir. Robert Mans and Dr. Ir. Rinke van Tatenhove-Pel, Associate Professors at the Industrial Microbiology Section, TU Delft
With Mr. Mans and Ms. van Tatenhove-Pel, we discussed other possible applications for our system. They thought that whole-cell biosensors could be a good implementation for our system. Another application could be bioethanol production in open tanks. GMOs are currently used in bioethanol processes but require closed production systems. For future developments, DOPL LOCK could be implemented in engineered organisms for use in open systems where it acts as an additional safety measure. Furthermore, an application of our system could be in wastewater treatment. These systems process large volumes and are generally operated as open systems, which limits the use of GMOs as they could escape the system via, for example, leakage, birds, rain and insects. In these systems, the induction of DOPL LOCK could for example be controlled by changing external conditions, such as temperature or pH, as Mans indicated that there are likely natural regulation mechanisms present in bacteria that respond to these factors.
Integration
Synthetic biology can be used for many different purposes. From our talks with Mr. Wagenveld, Mr. Gilbert, Prof. Ellis and Mr. Mans, we discovered that our proposed biocontainment system could have many applications. Even though Prof. Ellis mentioned that our system is likely to work , biocontainment systems are still not used. According to Mr. Gilbert, this was because of regulations, education and money. When developing our DOPL LOCK system, we need to take these reasons into account. Therefore, we did multiple educational activities and made a start-up plan on our Entrepreneurship page. Besides that biocontainment systems have not been used yet, we still wanted to identify the branch that could benefit most from a sufficient biocontainment system.
Our initial idea was to use our biocontainment system as a system to prevent the spread of antibiotic resistance genes. We thought antibiotic resistance genes were used to construct GMOs. After a meeting with Prof. Pronk, we discovered that the opposite was true. Using his feedback, we continued our search for the best application of our system.
Next, our team sat down to discuss other potential applications for our system. We concluded that the environmental biotechnology sector can probably benefit the most from our system. This thought was confirmed by Prof. Ellis, which stated that water and environment remediation applications and algal biofuel-producing cells grown in open water ponds are areas that need a proper biocontainment system. Bioremediation companies QM Environmental International BV and Micro-Bac confirmed that they could use a reliable and simple biocontainment system. With Mr. Wagenveld we talked about the PFAS issue , which can pose a risk to human health and the environment.
He emphasized that it would be very helpful to design a bacterial strain that could break down PFAS. Since PFAS is a seriously big problem, we decided to make a case-study about it. You can read about this case study on our Implementation page. In addition, Mr. Gilbert told us that in the future, the amount of non-degradable compounds like PFAS will rise. As a result, bacterial strains would have to be engineered to break down those compounds as there have not been successful alternative solutions yet. As a result, significant costs for the application and implementation of DOPL LOCK may be justified in these critical cases where GMOs are the only solution. Mr. Wagenveld and Mr. Gilbert from the environmental biotechnology field showed interest in our system. Another possible application could be in the field of whole-cell biosensors , according to Mr. Mans and Ms. van Tatenhove-Pel. We took this insight into our entrepreneurship plan.
We also took the three criteria for a good biocontainment system from prof. Ellis into account. Firstly we have designed our system in such a way that no complex elements (e.g. riboswitches or recombinases) are present in our design, which is primarily composed out of promoters, toxins and antitoxins. Additionally, DOPL LOCK can be designed in such a way that no special requirements (like medium composition) are necessary to apply the biocontainment strategy. Finally, due to the relatively small sequence length of toxin and antitoxin genes, our biocontainment system would be relatively mutation prone. However, we have explored additional strategies to make our system less mutation prone, which are discussed on the Safety page.
Possibilities within iGEM
Input
Tessa Alexanian, Safety and Security Program Officer at iGEM Foundation
Our idea was to make our system available for future iGEM teams to make their solutions safer. Therefore, we talked to Ms. Alexanian from the iGEM Foundation. Her hypothesis is that right now it is too complex to include biocontainment in the project from the beginning as (i) this would add an extra level of engineering and (ii) this would not be of importance for every team. If we could show other iGEM teams that it is easy to implement the system, it could be more appealing to use.
Integration
We envisioned the use of our system by other iGEM teams and think that previous iGEM projects could benefit from an easy-to-use and reliable biocontainment system. Below are three examples:
- HebrewU 2018: Catalysis of Dioxin degradation in the soil using GMOs.
- York 2014: Removal of Cadmium from wastewater.
- Wageningen 2014: Protect bananas from a TR4 plague.
It is hard to apply these three examples in practice unless they could implement a biocontainment system. After we met with Ms. Alexanian, our aim was to facilitate future iGEM teams to incorporate biosafety from the beginning of their project. We made our multiple cloning site biobrick compatible , which makes it easier to implement. You can read more about this on our Entrepreneurship page. With our iGEM project as the first step, we hope to create a world in which iGEM teams, startups, and industry can solve real-world problems using real science and real-world applications. Therefore we collaborated with iGEM Manipal biomachines, or MIT_MAHE, a team from India. We have worked closely with this team to design a combination of our two projects. We made a case-study about how we could implement DOPL LOCK into their project. For a more detailed description, we would like to refer you to the Implementation page.
-
Regulations
During the other sections of our Human Practices program, much discussion was focused on current regulations and possibilities for the semi-contained (applications in the environment where localized containment is possible, yet with a clear physical path for GMOs to spread into the environment) use of GMOs. Specifically, it was often said that the regulations are too strict and that they do not leave enough room for the development of GMO applications. For this reason, we also investigated what these regulatory restraints exactly entail and why these legislations apply.
Therefore, we spoke with regulation specialists and policy consultants to check whether the applications we had in mind for DOPL LOCK were realistic. With this, we wanted to formulate what room is left for semi-contained GMO applications and if this could change in the future. Furthermore, we also wanted to consider the perspective of the regulators and their views on the safety of GMOs.
Input
Cécile van der Vlugt, Senior risk assessor GMOs at the National Institute for Public Health and Environment & Marja Agterberg, Senior risk assessor GMOs and Project leader at Bureau GGO, National Institute for Public Health and Environment
Ms. Van der Vlugt and Ms. Agterberg informed us that the introduction of GMOs in the environment is possible, but that you need to apply for a license. The National Institute for Public Health and the Environment can help the applicant in understanding the needs in the risk assessment needed for this license. Van der Vlugt said that it could be an option in the future to take into account the benefits next to the risks and to work towards a risk-benefit analysis. This would ask for a change in regulation, however. Agterberg said that they have around 10 to 30 requests per year for the introduction of a GMO in the environment. Besides plant and clinical trials, work with optimized cyanobacteria outside of the lab was recently licensed [10]. We also talked to them about the reason why the European legislation is strict. They said that the societal acceptance of GMOs gives hardly room to review the regulation.
Rianne Hageman, Program Manager for technology for a sustainable future at HollandBio & Leonie White-Scholten, Project Manager for technology for a sustainable future at HollandBio
Ms. Hageman and Ms. White-Scholten mentioned that regulations are probably the main problem that prevents many GMO projects from being fulfilled. They think that EU regulations are a hindrance to future developments. Also, applying for a license requires a lot of administrative work. Smaller businesses and start-ups frequently lack the necessary funds and time. As a result, Hageman and White-Scholten believe that the regulations must be revised, as they were developed 20 years ago and are based on techniques developed 40 years ago. Besides regulations, acceptance is also a major barrier we need to overcome. According to them, some people oppose the use of GMOs. The legislation can be changed, but if nobody wants to buy it, the application will fail.
Clara Posthuma, Scientific Secretary of Medical & Veterinary Aspects at the Netherlands Commission on Genetic Modification (COGEM)
Posthuma indicates that there are worldwide strict regulations for the use of GMOs that scientists need to keep up with. She points out that the argument that there has never been a disaster, does not necessarily mean that GMOs are inherently safe to use. In fact, strict regulations might even have prevented big incidents. She mentioned that the current regulations may force scientists to think about safety and be more careful.
Marie-Louise Bilgin, Senior Policy Advisor Biotechnology at Ministerie van Infrastructuur en Waterstaat
The Dutch government agrees with the outcome of the study on New Genomic Techniques of the European Commission that current regulations are not fit for purpose. Current regulations will be reviewed in the future, but since the European Commission has to include all the European member states in the new propositions, this may take a long time. Every European country is bound by European regulations. Within the regulations, policymakers do have some leeway, but there is a limit. We also discussed the importance of risk analysis: it has to be shown that the risk of spreading and pathogenicity is minimal. Therefore, experimental evidence is required. The DOPL LOCK system of iGEM Leiden could lower the risk of spreading of a GMO and the risk of HGT.
European Food Safety Authority (EFSA)
We also contacted the European Food Safety Authority (EFSA) to discover how the European legislation regarding GMOs works. The environmental risk assessment (ERA), which is reviewed for GMOs by the EFSA, is a major component of current legislation. They mentioned that they are performing risk assessments for applications under the Directive 2001/18/EC [11]. This directive is the reference point for regulators and risk assessors. Any application for release into the environment will have to be compliant with this.
Britte Bouchaut, Ph.D. Candidate Safe-by-Design for inherently safe Biotechnologies at TU Delft
Bouchaut said that for the current regulations, an applicant needs to show that a process is safe. She mentioned that the potential risk with the semi-contained use of GMOs is that entire ecosystems can be affected. This is also the reason why the regulations are very strict. The policy is based on the precautionary principle: it is not permitted to introduce a GMO in the environment if it cannot be said with certainty that it is safe. Because this is never completely safe, we usually discuss acceptable risks. Lastly, Bouchaut mentioned that our system could be a valuable attribution to the Safe-by-Design principle.
Professor Jack Pronk, Full Professor and Head of Biotechnology Department, TU Delft
Professor Pronk thinks that, after over 40 years of safe use of genetically modified microorganisms in large-scale industrial processes, it is time to reconsider and relax some of the current regulations for their use. In particular, he is convinced that regulations on the application of microorganisms in the industry should be based on their properties rather than on the techniques used for strain improvement.
Integration
During our brainstorm phase, we already found that regulations regarding the non-contained use of GMOs are very strict. We wanted to know if all the applications we had identified were realistic. Also, we aimed to understand the reason behind the strict regulations and future developments in the legal system.
From our talk with Ms. van der Vlugt and Ms. Agterberg, we found that it is already possible to implement semi-contained GMO applications when allocated a license. Despite the difficulty of obtaining a license, a company in the Netherlands obtained one, demonstrating a successful application of GMOs outside of the laboratory. On the other hand, enough experimental data and knowledge on the legislation for this application are needed. Through our meeting with Ms. White-Scholten, we also discovered that the application for a license is a time-consuming job. We implemented this knowledge into our project by envisioning a company that facilitates such applications on behalf of other companies. For more information, we would like to refer you to our Entrepreneurship page.
From Ms. Van der Vlugt, we discovered that risk assessment is important for this license application. Although this risk assessment goes overboard according to some scientists, it is of great importance because it forces us to use GMOs safely , according to Ms. Posthuma. In addition, from our conversations with EFSA, we discovered that they used the Directive 2001/18/EC to perform this risk assessment. To make this directive more understandable and to show our consideration in these conditions, we made a regulatory roadmap that can be found on our Contribution page and used it to assess the risks of our project. We also show an example of our execution of this risk assessment within the case-studies on our Implementation page.
Via our conversation with Ms. Bouchaut we discovered that another important element of the legislation is the Safe-by-Design principle. Safe-by-Design entails incorporating safety into product and process development as early as possible. Its goal is to prevent environmental hazards and to provide a clean, healthy and safe living environment. The government's environmental policy includes Safe-by-Design [12]. Ms. Bouchaut emphasized the importance of using Safe-by-Design for application in the development of new technological processes. Therefore, we drafted a Safe-by-Design approach during the further development of our DOPL LOCK system. For example, the first step of this approach is the identification of the environmental conditions and risks , in order to subsequently tailor the DOPL LOCK system to overcome these identified risks. We intend to identify the environmental conditions and risks by following the steps outlined in our regulatory roadmap for environmental risk assessment (ERA). The tailoring is based on this risk assessment, which was concluded from the first step in the Safe-by-Design approach. To make this process of tailoring DOPL LOCK easier, we worked on a step-by-step approach of the plasmid design for our proposed implementation. For more information about the implementation of the Safe-by-Design principle, we would like to refer to the Entrepreneurship page.
Through our Human Practices, we discovered how the process of application for a license works, which will help us further along the line of developing our project into a company. In addition, from our talk with Ms. Hageman, Ms. White-Scholten and Ms. Van der Vlugt, we concluded that there is also a need for more social acceptance to enable a change in the regulations. This is why we delved further into this topic and conducted a variety of educational activities for various age groups at high schools and museums to explain the fundamentals of synthetic biology and its safety regulations, as described in the 'Social considerations' section below.
-
Social considerations
As you can read in our 'Regulations' section, some experts believe that the rules are strict due to public fear. To investigate this, we talked to social scientists and set up a survey.
Input
Britte Bouchaut, Ph.D. Candidate Safe-by-Design for inherently safe Biotechnologies at TU Delft
Bouchaut confirmed that the public opinion on GMOs is influenced by protests in the 1970s and 1980s. The main idea still is that GMOs are bad and unnatural. Another fear of people is that whole ecosystems can be influenced by the use of GMOs.
Kyra Delsing, Researcher Biotechnology at Rathenau Instituut
Ms. Delsing advised us on how to include stakeholder views in our research project. She explained different social science methods and mentioned that it is not easy to develop and analyze a survey when you are in the exploratory phase (not yet knowing who, what and how to ask the questions). In addition, she advised us to also engage in real-life conversations with stakeholders. This way you cannot have the same amount of participants, but you can be more flexible and ask follow-up questions to make sure you understand the views of the stakeholders.
Quotes from our survey participants
In collaboration with iGEM MIT_MAHE, we set up a survey about the public opinion on the release of GMOs. Below we show some answers on 'Can you describe the concerns and the objections that you personally have with the application of GMOs outside of the lab environment?' extracted from this survey:
'Outbreak of a species that behave in such a way we did not expect. I think we should always be ready to eliminate GMO's if not all things go to the plan'
'If it helps to fight diseases or climate change I am all for it, but by genetically modifying organisms they get altered too much, which might result in 'losing' the current 'type' of an organism. If they are protected from diseases and the risks are low, I do not object to'
'I would be most concerned with the way it interacts with the environments and other organisms in it. For instance, genetic modification of plants to enhance yield should not negatively impact insect populations. Proper research in a controlled environment should be performed'
'In a lab, it's a very controlled environment, where everybody involved knows the risks. If larger companies start working with it, they might seek financial gain over safety, and that could cause a lot of problems. In a lab environment, there are very strict rules that are followed. Companies could try and find loopholes, because they are less controlled, which would enhance the chance of something going wrong.'
'We don't know yet the impact on the ecological environment if it is not safe'
Integration
Social considerations should not be forgotten when talking about biocontainment systems and GMOs. Ms. Bouchaut told us that GMOs are still considered harmful and unnatural. When implementing our biocontainment system in the real world, gaining the trust of society is important. Therefore, we did multiple educational activities at high schools and museums for different age groups to explain the basis of synthetic biology and its safety regulations. More information about these activities can be found on the Public Engagement page.
The results of our survey have shown that people do have concerns about releasing GMOs into the environment. The best strategy when bringing our product to the market would be to make scientific literature more accessible and understandable to the general public. To this end, clear science communication should be written by scientists for multiple different news outlets and other sources with references to the easier scientific source. According to our survey, the public considers scientific literature (92%), books (44%), newspapers (32%), the government (28%), television (11%) and social media (9%) as trustworthy sources on the topic of GMOs. We integrated this by reaching out to multiple newspapers and a national radio station to collaboratively write news articles about our project, as you can read on our Featured page. These survey outcomes can also be useful for future iGEM teams: they can use it to inform the public about GMOs.
With the advice of Ms. Delsing, to also have real-life conversations , we decided to organize a discussion evening on GMOs. During this evening, we discussed the following three statements regarding GMOs:
- GMOs pose a real threat to humans, animals and/or the environment
- The regulations surrounding the application of GMOs should be more lenient
- It is ethical to make genetically modified humans
For more information about the discussion evening, we would like to refer to the Public Engagement page.
References
- Lee, J. W., Chan, C. T. Y., Slomovic, S., & Collins, J. J. (2018). Next-generation biocontainment systems for engineered organisms. Nature Chemical Biology, 14(6), 530–537. https://doi.org/10.1038/s41589-018-0056-x
- Anyshchenko, A. (2019). The Precautionary Principle in EU Regulation of GMOs: Socio-Economic Considerations and Ethical Implications of Biotechnology. Journal of Agricultural and Environmental Ethics, 32(5–6), 855–872. https://doi.org/10.1007/s10806-019-09802-2
- Martínez-García, E., Goñi-Moreno, A., Bartley, B., McLaughlin, J., Sánchez-Sampedro, L., Del Pozo, H. P., Hernández, C. P., Marletta, A. S., De Lucrezia, D., Sánchez-Fernández, G., Fraile, S., & de Lorenzo, V. (2020). SEVA 3.0: an update of the Standard European Vector Architecture for enabling portability of genetic constructs among diverse bacterial hosts. Nucleic acids research, 48(6), 3395. https://doi.org/10.1093/nar/gkaa114
- Rugbjerg, P., Dyerberg, A. S. B., Quainoo, S., Munck, C., & Sommer, M. O. A. (2021). Short and long-read ultra-deep sequencing profiles emerging heterogeneity across five platform Escherichia coli strains. Metabolic Engineering, 65, 197–206. https://doi.org/10.1016/j.ymben.2020.11.00
- Badran AH, Liu DR (2015) Development of potent in vivo mutagenesis plasmids with broad mutational spectra. Nature Communications, 6:8425. https://doi.org/10.1038/ncomms9425
- Valenzuela-Ortega, M., & French, C. (2021). Joint universal modular plasmids (JUMP): a flexible vector platform for synthetic biology. Synthetic Biology, 6(1). https://doi.org/10.1093/synbio/ysab003
- Novick, R. P. (1987). Plasmid incompatibility. Microbiological reviews, 51(4), 381-395.
- Sabri, S., Steen, J.A., Bongers, M. et al. Knock-in/Knock-out (KIKO) vectors for rapid integration of large DNA sequences, including whole metabolic pathways, onto the Escherichia coli chromosome at well-characterised loci. Microb Cell Fact 12, 60 (2013). https://doi.org/10.1186/1475-2859-12-60
- Yi, L., Chai, L., Xie, Y., Peng, Q., & Peng, Q. (2016). Isolation, identification and degradation performance of a PFOA-degrading strain. Genetics and Molecular Research, 15(2). https://doi.org/10.4238/gmr.15028043
- Stcrt. 2021, Nr. 36152. file:///Users/lisakleinjan/Downloads/stcrt-2021-36152.pdf
- 2001/18/EC of the European Parliament and of the Council of 12 March 2001 on the deliberate release into the environment of genetically modified organisms and repealing Council Directive 90/220/EEC. (OJ L 106, 17.4.2001, p. 1) http://eur-lex.europa.eu/legal-content/EN/TXT/?qid=1375683320071&uri=CELEX:32001L0018DIRECTIVE
- (n.d.). Safe-by-Design. Retrieved October 10, 2021, from https://www.safe-by-design-nl.nl/home+english/about+safe-by-design/default.aspx
