Team:Tufts/Safety

Safety

Commitment to Safety

Tufts Synthetic Biology promotes health, safety, and environmental protection in teaching, research, and public service by working closely with Tufts Environmental Health and Safety (TEHS) to ensure all members receive necessary training. TEHS provides leadership in developing and supporting high quality programs that allow our students to protect themselves from potential health hazards they could encounter. Tufts Synthetic Biology focuses on working with biological materials in a way that is safe for both researchers and the environment. In all cases, the researcher, lecturer, or third party is consciously aware of potential hazardous effects and has all necessary knowledge to do so safely, while meeting applicable laws and regulations.

Local Rules and Regulations

The project was completed on the Tufts University Medford campus, overseen by the Tufts Office of Environmental Health and Safety (Tufts EH & S) and the Institutional Biosafety Committee (IBC). The biosafety guidelines of our institution can be found at the following link: http://publicsafety.tufts.edu/ehs/biological-safety/ We submitted an IBC form to alert the University of the project we were pursuing, the location of our labs, and all the relevant details of how we would carry out the research. Our lab was inspected and the project was approved: IBC number - 2015-MR21 All of our team members have received and passed the Biosafety in Research Laboratories training.

Project Risks to the Researcher, Public and Environment

All bacterial strains used in lab are non-pathogenic E.coli. Work with Bacillus megaterium has not begun, but it is also BSL 1. The largest safety concerns our project created involved the intended delivery platform for CRISPR/Cas9. Our project thus far has involved the cloning of an atoxic Clostridium difficile toxin B, (atcdB) in DH5alpha cells (K12 strain bacteria). The toxin was rendered atoxic by mutations to catalytic amino acids in the toxin domain, as carried out and verified previously by Dr. Xingmin Sun at the Tufts Cummings School of Veterinary Medicine. The atoxic toxin was provided to us by Dr. Xingmin Sun. We have employed sterile techniques appropriate for a BSL-1 lab in cloning atcdB.

No work with the purified atoxic toxin, nor human cell lines, has yet occurred, however all work will be carried out under Biosafety Level 2 standards in a BSL2 laboratory, including restricting work to a biosafety cabinet. All work with human cell lines, including HeLa and HEK293T will be carried out under biosafety level 2 standards, in a BSL2 laboratory, including restricting work to a biosafety cabinet.

In addition to the atoxic toxin, we have plasmids in lab encoding resistance genes for ampicillin, chloramphenicol, kanamycin, tetracycline, and streptomycin. If our bacterial strains escaped, they could potentially transfer these resistance genes to other bacterial strains and species outside the laboratory. In order to reduce any risks associated with our project, we have employed sterile technique appropriate for a BSL-1 environment when working with our bacterial strains. Some chemicals that were handled in lab are capable of causing burns, irritation, allergic reaction, or death. Ethidium bromide and other intercalating dyes that were used in the lab have been shown to be powerful mutagens. Any potentially dangerous chemicals were handled in the fume hood using appropriate personal protective equipment (PPE). In addition, lab benches were wiped down with 70% ethanol before and after work while any waste was sterilized via bleaching or autoclaving.

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