Safety in Project Design

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During designing the project, we predicted and summarized any potential risks that will take place if the yeasts we used are being released. For the two substances 𝛼-pinene and isoamyl acetate used for attraction, if they are released, several consequences will occur and impact the environment:

1. Attraction to non-target creatures. Since we designed the 𝛼-pinene and isoamyl acetate for the attraction, there is a high probability that non-target creatures may be attracted to the surrounding environment. It will directly lead to the ecosystem imbalanced.
2. Genetic contamination. The released yeast containing 𝛼-pinene and isoamyl acetate could crossbreed with the natural yeast population potentially, which will result in the spreading of the genetic engineered genes which responsible for producing the 𝛼-pinene and isoamyl acetate and change yeast's internal genetic makeup of the original makeup inside the wild yeast.

There will also be negative results due to the releasing of the cry protein:

1. Negative effects on non-target insects. Though cry protein has high specificity for targeting and killing parasites and its harm to human beings is negligible, the non-target insects, especially moths and butterflies, will still be impacted.
2. The development of resistance to cry protein. If the cry protein is widespread and largely released, the nematodes we want to disinfect would develop its resistance to the cry protein we use.
3. Ecological impact. The leakage of the cry protein may affect the non-target organism. This could disrupt the balance of the ecosystem as beneficial insects, for instance, predators, may be impacted and lead to the increasing of the pests.

With the aim of preventing the possible danger, we design the kill switch system.

We design double insurance to ensure the yeast will not survive if it is released to the external environment. To ensure the kill switch can be carried out, we chose CEN.PK2-1C, which is an auxotrophic strain of S.cerevisiae as the chassis. The URA acts as the selectable marker, which implies that the yeast we used will only survive with the consistent supply of uracil in the closed medium we made.

As for the toxin-antitoxin system, Bax protein, and BI-1, the Bax inhibitor is mainly used for forming this system. Bax, the toxic protein, chiefly performs in mitochondria. The process of eliminating the cells could be briefly summarized by inducing the release of cytochrome c from mitochondria. The BI-1 is used to prohibit expression of the Bax protein and is located in intracellular membranes. In our closed system, the Bax protein is fused with constitutive promoter and the BI-1 is fused with inducible promoters. The amount of the BI-1 protein is larger than the amount of Bax protein, and the yeasts thereby survive. Once the yeasts are released to the external environment, the overexpression of BI-1 will immediately halt, so the yeasts won't survive, which could largely ensure biosafety.

Lab Safety:

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We also received the adequate training related to the organisms handling and any special experiments relevant from our instructor, Sha Zhou. The procedures of using different apparatus, rules for storage of different chemicals and usage of hazardous chemicals is inclusive in the training. Our instructor supervised us during the experiment to guarantee all processes are done precisely. A protocol including different procedures is also provided for helping us when we miss the procedures of the experiment. With the aims of ensuring team members better understand and memorize the rules relating to the lab safety, a test is also provided to examine team member's understanding.

In addition, we are asked to follow the rules strictly since we strived to conduct the all the experiments in the laboratory properly. For instance, according to the rules, eating is prohibited in the section we work in and all of the genetically modified organisms we use must be sterilized before they are released to the external environment of our laboratory. In addition, to ensure the personal safety of the team members, gloves, googles and lab coats are necessary before entering the experimental area as they could largely protect us from the harm of acidic or basic chemicals directly contact with our skin.

Safety promotion in human practice

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While basically ensuring the safety of our own projects and experiments to the outside world, we have received a lot of inspiration and help from the community. This inspired us to extend our sense of responsibility for synthetic biology safety from within the lab to the general public. Therefore, in our human practice activities, we place great emphasis on safety.

Privacy and respect in human subjects research

Safety is not only for our project and the third party, in human practice activities, it is also important to ensure the safety of interview subjects. This requirement forbids us to impose any form of negative impacts on our researched subjects. Hence respectful, ethical communications and value on the participants' privacy are essential. GreatBay-SCIE strictly adheres to the privacy protocol and ethics approval protocol in China. In every interview to ensure the privacy and security of the interviewees. Before each interview, the person in charge of the interview needs to outline the interview questions in advance after careful thoughts and research, in order to instruct a decent set of questions that is not only closely related to our project and problems, but most importantly would not discomfort the interview subjects. Then we send the questions to the interview subjects. The interview will be officially started when they have no dissidents on the interview content. At the same time, each interview question outline will contain our privacy agreement. After we fully understand the interviewee's acceptance of information disclosure, we will upload the interview content on the wiki page or completely anonymize it.

When conducting human practice activities such as in-street interviews, where it is impossible to inform the interview subjects in advance and discuss confidentiality issues, the team members will also ensure that the privacy agreement is shown to the interview subjects before conducting the interview, recording or filming. We would also practice the interview scenes ahead spontaneously to ensure that the interviewers hold polite and inoffensive manners.

Dissemination of biology lab safety

One of the starting points of all GreatBay-SCIE education activities is to promote the safety of biological laboratories, and we have implemented safety in the majority of our education activities.

1. Lab sessions are set up in ECA on campus
   
   

   The team members agree that the significance of ECA is not only to simply promote the knowledge of synthetic biology, but also to ensure the safety of practice in this experiment-based subject. Therefore, we specially included lab session in this year's ECA course, aiming to let students experience the charm of synthetic biology in person, and cultivate their attention and understanding of safety matters in the laboratory.

   

   Figure 6. lab session in ECA under discreet monitor by team members to ensure lab safety.

   In each lab session, the team member in charge of this experiment teaching must include risk assessments and corresponding safety measurements in the compilation of the experiment protocol. After passing the team’s approval, Will be distributed to community members in the lab session.

   

   Figure 7: lab safety precautions provided in the experiment protocol, which was handed out to every participant for guidance.

2. Upload experiment log on online social platform (RED)
   

   When the project is progressing, the team members will also regularly record the process of some common experiments (such as PCR) by video and upload it to the social platform, aiming to publicize synthetic biology and demonstrate the correct experimental operation mode and experimental safety matters that must be observed to those interested in biological experiments, so as to raise people's awareness of experimental safety.

   

   Figure 8. account of GreatBay-SCIE on social platform RED. Content covering basic synthetic biology experiment protocols and safety.

Proposal:

With the development of the synthetic biology, more and more synthetic biology related laboratory now occur in Shenzhen. However, we identified the shortcomings of the laboratory management regulation. There was no universal legislation or regulation rule for the commercial laboratories or medical and research level laboratories and a large amount of the potential risks could therefore being exposed. We thereby had the idea of writing a proposal, which is the collation of the legislations and rules for management regulation. Nevertheless, after the interview with Dr. Zheng, he gave us some advice about our thoughts and we later decided to write a suggestion about conducting a website which summarizes all the legislation and rules relevant to the management regulation of the laboratory under his advice and instruction. The main purpose of this proposal is to provide a better and more convenient way for experimenters searching for relevant legislation and rules and carrying out safe experiments. It can also be useful for regulating the management for the laboratories as most legislation and rules are complex and hard to find out and,with this platform, relevant personnel who in charge of managing the laboratory can find out this with shorter time. The new proposal mainly contains several suggestions:

1. The platform, which is the website, should contain the relevant management regulations. However, all of these rules should be classified into different categories, because laboratories at different levels have different management regulations. For instance, we can divided the platform into two categories, one for enterprise level and other one for medical level, which for better references.
2. All the relevant regulations of Food and Drug Administration, and National Health Commission and other rules of municipal and provincial level should all be included. As they should all be followed.
3. Besides, for the enterprise level laboratories, those related to the education especially, should summarize all the verbal rules and regulations into text form for better education. As for better regulating these rules, a seperate area can be used to record these verbal rules. In addition, these regulations should also be disseminated among the laboratories in order to better refine the oral regulations. Experimenters and especially the supervisors in the laboratories, should also give these files to the students and ensure they know all of this which will ensure their safety.
4. Details about the platform

A menu is designed for better accessing different legislations and regulation rules.

We basically summarized several aspects as the menu of the webpage.

The webpage should be mainly separated into two parts, which are Medical and research level laboratory management and commercial laboratory management. And below we made an example of the menu, and some example legislations and rules are also provided:

(Click Here for Downloading the PDF Document)

The proposal is handed in to the local government and they recently gave us positive feedback, which is our proposal has a high probability could be used in the upcoming year, even could be used in 2024.