Our lab, an extension of the Lynch lab, follows the BSL-1 guidelines described as suitable for work involving well-characterized agents that are not known to consistently cause disease in immunocompetent adult humans. The guidelines include washing hands before leaving the laboratory, and careful management of needles and other sharps. Decontamination is strictly followed through autoclaving of solid trash, bleaching of cell waste solutions, and separately disposing of special waste, such as chloramphenicol. In terms of safety equipment, gloves are worn at all times and changed when contaminated. More details on the guidelines can be found at the CDC page and the Duke Safety page.
We received training on general laboratory safety, including personal protective equipment, proper handling of biological and hazardous waste, proper handling of sharps, use of fume hoods, and emergency procedures. We were also trained on general chemical safety, including identification and labeling of hazards, proper storage and use of common chemicals, regulations regarding more hazardous substances, waste disposal procedures, and basic first aid and emergency procedures. In addition, we also learned about fire and life safety, including emergency preparedness and policies.
This training was through various forms of online and in-person training, required by Duke, depending on the safety levels and hazards you are expected to face. For our purposes, we are required to complete online courses for General Laboratory Safety, General Chemical Safety, and Fire/Life Safety. These courses include online modules and a post-course quiz. A description of required training and the training policy is provided at the Duke Safety page.
Local Rules and Regulations
The Occupational and Environmental Safety Office of Duke University covers safety concerns and compliance. We are working directly within the Lynch lab, which are compliant with and in communication with this office, and we have completed the required safety training that they provide. The guidelines are as described in the Duke Biological Safety page.
Organisms and Parts Used
The chassis organisms used for cloning are the E. cloni strain of Escherichia coli by Lucigen, which are in risk group 1 . They were acquired from the Lynch lab.
Duke iGEM's goal is to optimize the biosynthesis of the chemotherapy drug taxol in E. coli. Our objective is to individually characterize and then consolidate known enzymes of the taxol biosynthesis pathway into a single strain of E. coli. As such, our research plan consists primarily of two components: a cloning project and kinetic assays to characterize the system. Our strain of E. coli would be used in fermentations to synthesize taxol.
Laboratory Risk Factors and Mitigation
The biggest risk we work with is SYBR Stain, which is a mutagen and irritant, although we use it in small concentrations. Another risk is the antibiotic chloramphenicol, which is strongly anticipated to be a carcinogen by the National Toxicity Program . The E. coli that we use are derived from a K-12 strain and therefore are not particularly dangerous to humans. However, contact with bacteria could still lead to infection or illness, particularly if the bacteria are ingested. In our lab, we wear gloves, sterilize waste, and keep chemicals in cupboards.
Future Risks and Applications
The project is harmless to average real people because the product is a manufactured chemotherapy drug, meant to be regulated and administrated by the health care industry. However, there may be some issues concerning the purity of the taxol derived from E. coli, which would require further precautions.