Team:RDFZ-China/Safety

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Safety

Introduction

In iGEM, safety covers the procedures, practices, or other measures used to manage risks from accidental exposure or release. Security covers the procedures, practices, or other measures used to manage risks from deliberate exposure or release. Working safely and securely is a core element of responsible research and innovation. RDFZ iGEM team addresses safety and security issues throughout the competition lifecycle. Team members consider potential risks to themselves, their communities, and the environment, and try to manage any risks through the process of project design, laboratory work, and human practice.

1. Safety and Security in Project Design

1.1 Organisms Used

The E. coli strains we use are whitelist species and are a communal strain that has been used as a probiotic agent to treat gastrointestinal infections in humans since the early 1920s. If the E.coli is released into the environment, due to the weakening of its varieties, its survival ability has been greatly reduced, which will not cause ecological problems. Occasionally, however, probiotics may have adverse clinical effects, including septicemia. If both the microbiota and adaptive immunity are defective, translocation across the intestinal epithelium and dissemination of the probiotic E. coli strain may occur. Considering this risk, we plan to use Escherichia coli Nissle (EcN), a non-pathogenic probiotic strain isolated in 1917 by Alfred Nissle, in practice. E.coli Nissle 1917 has been commercially available for almost one century with no toxicity, and it is one of the best-studied probiotic bacterial strains. Therefore, our bacteria are not harmful to the environment or the community.

1.2 Experiment Design

We will conduct Gibson assembly on our plasmids, transform human TPH1 enzyme into E.coli Nissle, use a fluorescence assay for Tryptophan Hydroxylase to test its activity, and conduct experiments to measure the sensitivity of our circuit. Since we decided to do a two-year project, the first year will only focus on hardware design, plasmid construction, and predicting experiment result using modeling. Since this year we are not going to do stimulation of our bacteria hardware in mimic human body environment, there are fewer risks of bacteria leakage except the occurrence of mild discomfort or diarrhea when contacting with E.coli.

2.Safety and Security in Laboratory Work

2.1 Training

enough to equip team members who participate in wet-lab experiments with necessary knowledge and skills that ensure biosafety and biosecurity. The supervisor offers a lab instruction booklet and an online quiz to make sure every team member knows the basic lab operation. All team members are required to complete the rigorous examination related to biosafety and biosecurity before conducting wet-lab experiments. The exam contains 42 questions and covers every aspect of the lab activity. Members who fail to score over 80 percent are forbidden to enter our labs. We learn about lab access and rules (including appropriate clothing, eating, and drinking), responsible individuals (such as lab or departmental specialists or institutional biosafety officer), differences between biosafety levels, biosafety equipment (such as biosafety cabinets), good microbial technique (such as lab practices), disinfection and sterilization, emergency procedures, transport rules, physical biosecurity, personnel biosecurity, dual-use and experiments of concern, data biosecurity, and chemicals, fire, and electrical safety.

2.2 Lab Setting

The biosafety level of our workplace is level 1. The labs in which all our wet-lab experiments are conducted were built under the supervision of experts and were approved by the Chinese government. Risk management tools including accident reporting (measures to record any accidents), personal protective equipment (including lab coats, gloves, eye protection, etc), inventory control system (measures to track who has what materials and where they are), access controls (measures to control who can access your workspaces, or where materials are kept), waste management system (measures to make sure waste is not hazardous before it leaves your institution) cover our work. The officers provide all assistance in experiments requested by our members such as the usage of PCR machines, cleaning used instruments, and turning off the electricity and flames before leaving. Our supervisor researched a lot of the usage of our E.coli, and he is familiar with the experimental procedures and practices.

2.3 Containment

When conducting wet-lab experiments, all team members are supervised by experienced lab experts, and the protocols used are seriously examined to ensure biosafety and biosecurity. All team members are required to dispose of waste appropriately. They strictly follow lab protocols and make every effort to prevent leaking.

3. Safety and Security in Human Practice

3.1 Surveys and Interviews

We engage with communities to understand broader social concerns by talking with specialists in this area. All human subjects research conducted by teams is done in accordance with relevant laws and regulations, as well as any institutional rules or guidance, and above and beyond in demonstrating the commitments to responsible and ethical conduct. Our human subjects research is ethical and responsible for issues such as informed consent and privacy and data protection. Recorded verbal consent is made and specific permission from the cited individual to use their name has been obtained.

3.2 Real Application

If our project was fully developed into a real product that real people could use, it might have adverse clinical effects, including septicemia. If both the microbiota and adaptive immunity are defective, translocation across the intestinal epithelium and dissemination of the probiotic E. coli strain Nissle 1917 may occur and have potentially severe adverse effects. If TPH1 level were high enough, it might produce too much 5-HTP, resulting in too much serotonin, causing serotonin syndrome, which might have severe health effects. We eliminate the risk by adding a strict negative feedback loop.