Brainstorming with instructors

When our team was first established to participate iGEM, we were distressed at what a product we can produce by synthetic biology. So our team began to read the previous award-winning work of iGEM so as to get some inspiration for our project. But in pace with our reference work, we found that there was a growing awareness on the importance of a safe chassis organism for the work of synthetic biology, as well as potential environmental and health consequences of developing and using synthetic biology technologies and products. For example, we are very concerned with increasing projects of synthetic biology all over the world will definitely increase the chance for lab leakage of the genetic engineered chassis organisms, which may then bring risks of our environments and human health. Meanwhile, we also found that The World Health Organization (WHO) released Global guidance framework for the responsible use of the life sciences (2022), proposed that scientists are the first line of control for assessing, preventing and mitigating risks, and they have a professional responsibility to consider and mitigate risks from knowledge, products or technologies that they develop and disseminate used for harmful purposes.

Therefore, our team came up with an idea that we will develop a more secure chassis platform of SynBio, which would be very hard to accumulate out of laboratory conditions. We believe this type of biosafety-focused research is certainly worth more attention from SynBio research. If successful, the new platform can be promoted to utilization of various microbial chassis and application worldwide, the risks of lab leakage could be significantly reduced. Meanwhile, within this project, we plan to learn about the regulation and law related to safety governance of synthetic biology, and then advocate the importance of “Responsible Synthetic Biology” for high schoolers and iGEMers, as they are the young and future generation of synthetic biology researchers.

Figure 1. Our team members were brainstorming about our project and practiced lab skills for cyanobacteria cultivation and transgenic engineering in the biolab of Tsinghua International School Daoxiang Lake (THISDL).

We the initiated a discussion and solicited suggestions from Prof. Weiwen Zhang at Tianjin University Chemical Engineering School and the Center for Biosafety Research and Strategy about our idea and got very positive reply early January 2024. Prof Zhang gave us very benefiting lectures about the policies and techniques for biosafety related to synthetic biology and encouraged us to conduct experimental work in the Synthetic Microbiology lab at Tianjin University. Before we started our lab work at Tianjin University in the early summer of 2024, we also got further instructions and training for relevant techniques and lab skills from Prof. Lei Chen. Prof. Chen recommended us some critical references about biocontainment technology and suggested us to use photosynthetic cyanobacteria as the model platform to develop our techniques, since cyanobacteria has been widely used as synthetic biology platform in recent decades due to its capability to utilize sunlight and CO₂ as the sole energy and carbon sources, respectively; meanwhile, the high survival ability of autotrophic cyanobacteria in nature could bring a higher risk for the environments compared to other heterotrophic chassis. At our request, Prof. Chen sent us some cyanobacteria samples and plasmids from his lab to Tsinghua International School Daoxiang Lake, which gave us the valuable opportunity to practice some key lab skills about cyanobacteria cultivation and genetic transformation, even before to come to Tianjin University Laboratory in the summer.

Figure 2. Our team discussed with Prof. Weiwen Zhang, Prof. Lei Chen and Prof. Kun Zhao, as well as graduate students at Tianjin University, about the feasible technology roadmap for our designed biocontainment system.

With the help from Prof. Zhang and Prof. Chen, especially the papers they recommended to read about biocontainment, our team finally found a promising route for our technique development, namely a temperature-sensing biocontainment system, because this system does not need to add any exogenous chemical inducers, reducing any secondary chemical contamination to natural environments, and the temperature-controlled gene expression can be simply accomplished by the temperature-sensitive promoter P_R (corresponding repressor cI857) detected in bacteriophage. In terms of technics of how to kill the escaping cyanobacteria, from reading papers, we found that there was a native endogenous toxin-antitoxin system in the cyanobacteria strain Synechocystis sp. PCC 6803. Then, we realized that, if we could express the antitoxin gene under the control of the PR promoter, a logic gate based killing system controlled by temperature could be developed. The only defect of system was that the toxin-killing requires several days’ time, during which the escaping bacteria still have chance to reproduce and even exchange DNA with other similar microorganisms in the environment.

Therefore, we further turned to helps from Prof. Kun Zhao, who is an expert on bacterial mobility and microscopy. Prof. Zhao suggested that we could at first restrict the mobility of the engineered cyanobacteria before they were killed by the toxin and this system can be accomplished by destroying their pili. Then, we studied the literature and came up with an idea that CRISPR-dCas12a technique can be used to knock out pili-relevant genes, while the expression of the the crRNA against the target gene can also be controlled by the temperature-sensing P_R-cI857 system. If this double-safe system could be constructed, the risk of chassis organism leakage would be significantly reduced and this biocontainment system would in principle be adaptive to many other microbial chassis systems used by many other SynBio labs around the world.

With such ideas and expectations, our teams entered Prof. Chen and Prof. Zhang’s lab at Tianjin University to start our work on wet lab and dry lab with full of challenges and wishes.

Discussion with law experts about biosafety

In order to integrate our technique to the solution of technical, regulation and law. Our team invited Prof. Yang Xue at Tianjin University Law School and the Center for Biosafety Research and Strategy, who is the participant to draft the Tianjin Biosecurity Guidelines for Codes of Conduct for Scientists, to give us a lecture. In this lecture, Prof. Xue the introduced the potential benefits and associated risks of synthetic biology research, and emphasized the need for a balanced managemental strategy. He briefed us that since its formal start in early 2000s, synthetic biology has grown rapidly, driven by increased global investment and interest. This emerging field combines biology and engineering to design and construct new biological systems with applications ranging from novel medical therapies to sustainable biofuels. However, the rapid growth of synthetic biology also brings with it significant risks that need to be carefully managed.

A major concern is the misuse of synthetic biology. The accessibility of advanced tools and knowledge increases the risk of unauthorized applications. Policymakers and researchers are particularly concerned about dual-use of synthetic biology, as it could be mistakenly used to generate harms to environment and health, or in the worst cases, intentionally abused by bad actors to create harmful biological agents or toxins against human being. If synthetic biology research are not responsibly conducted and governed under the existing regulatory frameworks, any violations, whether intentional or due to inadequate regulation, can cause serious consequences.

China's regulation of biotechnology began in 1993 with the Measures for the Administration of Genetic Engineering Safety and has continued to evolve through a variety of regulations, including those governing laboratory biosafety of genetically modified organisms (GMOs) and pathogenic microorganisms. These regulations aim to manage risks and ensure safe operations in the field.

Through literature study, the team also found many published articles discussion on the potential benefits and risks of using cyanobacteria as “green cell factories” because of their ability to use solar energy to convert carbon dioxide into organic matter. Advances in biotechnology, such as synthetic biology and genome editing, are enhancing the use of microalgae in medicine, agriculture, food production, energy and environmental management. However, engineering microalgae also poses biosafety risks, including potential negative impacts on the environment and threats to human health. To mitigate these risks, it is critical to develop biosafety systems. The systems developed previously include active strategies (e.g., designing kill switches using toxic proteins) and passive strategies (e.g., creating nutrient-deficient strains by knocking out important genes). They are designed to control engineered microalgae and prevent unintended environmental spread.

After reviewing the recent advances in microalgae bioengineering, associated biosafety risks, and progress made in developing biocontainment technologies, the team concluded the importance of a balanced approach to harnessing the benefits of synthetic biology while responsibly managing its risks.

At the lecture on synthetic biology biosafety by Prof. Xue, the team sensed a sense of urgency in his words, also concerned about the biosafety risks mentioned. They asked teachers to clarify what risks engineered cyanobacteria poses to the environment and to human health, and learned how the engineered strains could damage local ecosystems if they escaped from the laboratory. The team also followed up with a question about strategies to ensure biosafety. The instructor detailed both active and passive strategies, including the intriguing concept of “lethal switches.” The team was fascinated by how these mechanisms could help control engineered organisms.

When the topic turned to regulation, the team seized the opportunity to ask how regulation plays a role in risk management. My teacher emphasized the importance of established frameworks like China's in overseeing research practices. The team was reassured to know that China has some systems in place to minimize potential harm.

From the lecture and literature studying, the team gradually realized that researchers are at the frontier of the synthetic biology innovation and should also be the first line of defense against misuse or abuse of synthetic biology technology. The team studied the "Tianjin Biosecurity guidelines for scientists” and concluded that some level of consensus on self-disciplining and the responsibilities of scientists conducting synthetic biology research are important among the synthetic biology community. For example, an awareness of the possible dangerous consequences associated with the research, reporting or terminating research when any sign of danger occurs, and barring publication of results possibly related to nefarious application. Meanwhile, the team also realized that no enough attention has been given to this important challenge among young high school students and iGEM contesters, so they decided to make a poster from what they learned to advocate the concept of “responsible synthetic biology research” .

Participating the CCiC 2024

In July12-15, our time went to participate the 11th Conference of China iGEMer Community in Suzhou, China, which last about three days long. During this period of time, we saw and learned different groups’ objections and their technique to achieve their goals. Our teams had learned a lot during viewing others presentation.

Later, we made our own presentation, which is about our newly investigated biocontainment system for cyanobacteria, and answer question from the judges in this forum.

Many other teams were interested in our team’s subject, and we had a close discussion with them because they have a similar objective and we looked forward for future cooperations.

During the conference, the team got the chance to present its posters on responsible science and relevant law and regulation in China. The team also delivered print-out of the poster to more than 100 attenders of the conference.

During the interval of the meeting, we were fortunately to have a project discussion with iGEM coordinator Mr. Yuhan Bao, Judge Mr. Shan Jiang and Prof. Hanzhi Yu from Zhejiang University on further improving our project. They made many valuable practical suggestions .

This experience in the forum helps the team to build a solid foundation for our project, as we learned the bright point and the deficiency in the project which we can improve in the future.

One of our team member interviewed professor Bhagwati Gupta from McMaster University (Video 1). They discussed the current development of synthetic biology in North America. Professor Gupta introduced his biggest concern about synthetic biology and the policies made by Canada and US to let relevant projects be limited and well controlled. He also advised us to definitely do experiments many times to make sure the temperature-controlled switch will turn on and off ideally. This advice is really valuable and will undoubtedly help refine and enhance the effectiveness of our project.

Video 1. The interview video with Prof. Gupta

Visiting water treatment facility

Our team went to a water treatment facility in Lang fang Hebei to learn about the steps of cleaning drinking water have a deeper understanding about the importance of the project we are currently working on.

In the facility, the team studied about the steps of doing the water cleaning work for example the sedimentation and the filtration. The first thing to remove from the water is the soil, earth and mud, the solid that can be easily seen by human’s eyes. The facility uses a pool to let these things become sedimentation at the bottom of the pool and then followed by uniform pumping. After that, they use chemicals to remove the bacteria and toxins from the water.

It is well known that cyanobacteria toxin is harmful for creature for example human. Cyanobacteria create several types of toxins, that will cause creatures diseases or even death. These toxins are called microcystin, that meanly including hepatotoxin – which cause harm to the liver, and is the most common toxin in the cyanobacteria, endotoxin-not that common but can cause death, and neurotoxin-which is the most toxic one among these three. There are 46 types of cyanobacteria that we already known it is toxic, and according to the report from WHO, 59% of the microorganism that cause the blooming are toxic cyanobacteria. This kind of cyanobacteria is harmful to the environment, here are two examples.

In the summer of 2007, about 2 million residents in Wuxi, China were left without drinking water for more than a week because Tai Hu, one of the biggest lake in China that is an important resources of drinking water for Chinese people, enveloped with a massive cyanobacterial bloom. In Aug. 2014, the city of Toledo, Ohio issued a "Do Not Drink" advisory for its residents when Lake Erie was covered with a thick blanket of cyanobacteria. On Aug. 6, 2021, Spokane, Wash. reported the death of 3 dogs barely 20 minutes after they went swimming in water contaminated with toxigenic cyanobacteria (from Cyanobacteria Blankets of Doom: Causes and Effects of Toxic Blooms, asm.org).

These toxins cause problems to life not only human, but also wild animals. A news from CNN posted on 2nd July 2020 said that in the 3 months from March of 2020 to the July of 2020, over 350 elephants dead in Botswana, that is unusual. 70% of them dead near to the water. “One elephant had been observed a behavior of walking in circle, unable to change direction although being encouraged by other herd members” said the report.

It is not easy to remove these toxins from water, so it will be important to avoid the water blooming. And this is what our project did, stopping the engineered cyanobacteria with growth or other physiological advantages escape from the lab can avoid some of the water blooming and protect our environment. The activity strengthened our idea about the importance of cyanobacteria anti-leakage system and encouraged us to improve our technique to play its social value soon.

Science popularization lecture to peer high schoolers

After we completed our experiments, our team organized lectures at different secondary schools in Beijing to introduce the importance of biosafety and the results of our project to the peer high schoolers of our team members. In order to bring more interest in our lectures, we also designed some interaction games with the audience. The participants in these lectures gave us very positive feedback. Most of them told us that they knew the mechanisms behind the risks of biosafety for the first time and the so fascinated with the synthetic biology techniques. Some students developed very high interest in synthetic biology after listening to our lectures. All these activities and feedback made us more confident with our technic and advocating efforts. We hope that we can continue working and optimizing our biocontainment system in the coming years. We also promise ourselves that we will practice our slogan “Science for Nature and Humanity” throughout in the future research.

Responsible Synthetic Biology for Nature and Humanity – Action from Youth

During the 2024 iGEM Grand Jamboree in Paris, the team plan to present a poster and statement entitled as “Responsible Synthetic Biology for Nature and Humanity – Action from Youth”. The team will re-state that scientists are at the frontier of synthetic biology innovation, and also the first line of defense against technology misuse and abuse. Therefore, to prevent the misuse and abuse of dual-use synthetic biology, it is essential to enhance the awareness and engagement of scientific community on biosafety issues and to reinforce the moral self-discipline of the scientific community. The team will introduce to all iGEM contesters (young generation scientists) several initiatives that have articulated ethical principles and toolkits to harness the power of science and innovation, including WHO’s 2022 Global Guidance Framework for the Responsible Use of the Life Sciences highlighted the concept that scientists constitute the first line of control for assessing, preventing, and mitigating risks, explaining that they have a professional responsibility to consider and mitigate risks that stem from the knowledge, products, or technologies that they develop; and the Tianjin Biosecurity Guidelines for Codes for Scientists that also identified a set of 10 guiding principles and standards of conduct for scientists to promote responsible science practice and strengthen biosecurity governance at various levels.