Team:CSU Fort Collins/Safety

Safety Considerations

Safe Project Design

Adriana all geared up to extract trans-zeatin

Except for our kill-switch, our parts are adapted from native genes in plants and bacteria. Our particular genes do not harm any type of living organism. Our breakdown part is designed to help the cells flourish. The trans-zeatin production construct makes a molecule that is beneficial to plants, and harmless to humans and bacteria.

As a manufacturing track team, we wanted to create something that could be feasibly scaled up. When discussing scale up into industrial size, the potential for an accidental spill becomes inevitable. Our product has to be either quick and easy to clean up or nonviable once released from our reactors. We decided to go the nonviable route. In addition to using a laboratory strain of E. coli (DH5alpha) as our chassis we included a kill switch in our design. The idea was to suppress the kill switch gene while the cells were contained and if they left that containment the cells would die. Through our research we found that our use of a kill switch for cellular level bio containment was insufficient. For more information on our characterization of KillerRed, see our results.

Safe Lab Work

All CSU iGEM members are trained in various safety protocols. Before getting access to the lab every member had to be certified in BSL 1 safety procedures, hazardous waste creation and disposal, and undergo an orientation on the building we work in and the specific equipment in it. Standard safety measures such as long pants, gloves, goggles, and biosafety cabinets were used every day to protect both us and our organisms.

Our lab work mostly involved chemicals typical to synthetic biology. For some of our experiments, we did have to take extra safety measures to protect ourselves. To test our breakdown constructs, we wanted to grow our E. coli in a media where the only carbon source was fatty acids. Fatty acids are very hydrophobic and suspending them into a media that cells can grow in requires surfactant (detergent). The first papers we found that grew E. coli on fatty acids used the surfactant Tergitol NP-40. This is a strongly corrosive detergent, and would require face shields to work with. Out of concern for safety, we continued our search for a surfactant that had been proven to work in literature and had fewer hazards. We eventually found Brij 58, which is very safe to work with.

For our work in extracting and identifying trans-zeatin, we also used extra caution. We were required to work with high concentrations of acetic acid, methanol, and formic acid during our extraction (SPE) and detection (HPLC). We used safety glasses, lab coats, and gloves in conjunction with a fume hood to ensure our safety.