Team:Nevada/Safety

1. Would any of your project ideas raise safety issues in terms of research, public or environmental safety? Our project is focused on developing a self-contained culture system in which a photoautotrophic organism (i.e. Synechocystis PCC 6803) would provide carbon nutrition to an ethanol or fatty acid secreting Escherichia coli strain. The one public safety concern with this year’s project was the BioBrick part that produces ethanol. Since ethanol is a control substance, we contacted the University of Nevada Environmental Health and Safety Department to inquire about the issue. The University of Nevada officials granted us permission to execute the project with the following restrictions:
• 1) large volumes of ethanol cannot be produced
• 2) the resulting culture medium will not be distilled to purify the ethanol
• 3) consumption of the ethanol containing culture medium or any of its derivatives is strictly prohibited.

To prevent any environmental safety concerns, we have taken precautions to prevent the ability of our experimental organisms from being able to survive outside of our controlled laboratory environment. In regards to our E. coli strains, we are working with the NEB10 Beta strain of E. coli (New England Bio Labs) that is deficient in producing leucine and cannot survive on media that is not supplemented with this amino acid (auxotrophic mutant).In regards to Synechocystis PCC6803, we have chosen to create our own auxotrophic mutant by using homologous recombination to create an insertional mutation in the thiamine monophosphate pyrophosphorylase synthase (ThiE) gene. In doing so, we will be creating an auxotrophic mutant that requires thiamine to grow and cannot survive outside the laboratory. Also, we are currently designing a self-contained apparatus that will be used when co-culturing our E. coli and cyanobacteria. This apparatus will prevent introduction of organisms into the environment and can be autoclaved. 2. Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues? The majority of the genes we are using in this project have been obtained from the iGEM parts depository or other iGEM researchers. As far as we are aware, these genes have not demonstrated any health or environmental risks. We have also generated new parts related to fatty acid production in E. coli and hexose sugar production in Synechocystis. Based on our search of the literature none of these genes nor the products of the encoded proteins pose any public, environmental or research safety concerns. The only concern relates to a BioBrick part that produces ethanol. As we described in Section 1, we have taken appropriate action to gain institutional approval for the use of this gene and are taking appropriate safeguards to protect the public, our researchers and the environment. 3. Is there a local biosafety group, committee, or review board at your institution? In order to prepare our team members for lab work, there are two safety training courses that are required for all members to attend. The first course is a basic safety class outlining the proper way to handle and dispose of chemicals and biological hazards. The second safety course is to teach us the ethical and safety issues associated with genetically engineering organisms. On our campus, all labs are required to adhere to rules and regulation set forth by the Department of Environmental Health and Safety, as well as the National Institutes of Health (NIH) for recombinant DNA. 4. Do you have any other ideas of how to deal with safety issues that could be useful for future iGEM competitions? How could parts, devices and systems be made even safer through biosafety engineering? For any iGEM team working with cyanobacteria, we would highly recommend that they make their own auxotrophic mutant by undergoing homologous recombination at the ThiE locus or other known location on the chromosome that would result in the production of an auxotroph.