Team:Michigan/Safety
Safety
With safe laboratory practices in mind, we designed a CRISPR Cas9 Testing Model that could help the efficiency of developing modifications of Cas9.
All team members underwent the University of Michigan’s Department of Occupational Safety and Environmental Health safety training for basic wet lab work as well as specific training for safe autoclave usage, and reviewed our lab space and practices. They concluded that our research was conducted in a way that minimized risk.
We used E. coli as our chassis organism because it is widely used in synthetic biology and easy to work with. It is a Risk 1 type organism, which means it poses little to no threat to human safety. Ampicillin, Kanamycin, and Chloramphenicol resistance was included as selection factors for our three plasmids during our experiments. We prevented the spread of this resistance with careful containment and sanitation.
Cas9, the main component of our project, specifically targets specific DNA fragments based on the guide sequence used (Gasiunas et al, 2012) and without a guid sequence available, Cas9 is unable to bind to DNA and nucleate it. The concentrations produced in this project poses little danger to eukaryotes.
Gasiunas, Giedrius, et al. Proceedings of the National Academy of Sciences of the United States of America, National Academy of Sciences, 25 Sept. 2012, www.ncbi.nlm.nih.gov/pmc/articles/PMC3465414/.