iGEM Safety Questions:

1. Would any of your project ideas raise safety issues in terms of: researcher safety, public safety, or environmental safety?

All Organisms used are BSL1 rated. The ''Escherichia coli'' strains used are were DH10B, DH5α, and MG1655. These ''E.coli'' strains have been engineered for lab use and contain no pathogenic factors. Wild type strains would most likely outcompete them in the event of an environmental breach. Additionally, the recombinant parts that they contain would likely strain the metabolic pathways of the organism, furthering the wild type E. coli’s capacity to outcompete them. The plasmids introduced into these strains only contain antibiotic resistances standard to the parts registry, including: ampicillin, kanamycin, tetracycline, chloramphenicol, and streptomycin. These do not otherwise grant the organism unique advantages/characteristics over their native brethren. As with all recombinant DNA, our project should not, without proper extensive testing, be exposed to the environment or public. If this were to occur, none of our recombinant strains should pose an imminent threat to environmental or public health.

2. Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues? If yes, did you document these issues in the Registry? how did you manage to handle the safety issue? How could other teams learn from your experience?

We will be submitting two main parts to the registry this year. The first is a translational coupling cassette, which contains a red fluorescent protein and an engineered mRNA hairpin. This should not pose any safety risks; it is a diagnostic tool, and poses no risk to public health/safety. The second is a limonene synthase part. This allows for the production of limonene inside the cell, but since limonene has only been observed to be a respiratory irritant after long term exposure, this should not pose any human health risk.

3. Is there a local biosafety group, committee, or review board at your institution?: If yes, what does your local biosafety group think about your project? If no, which specific biosafety rules or guidelines do you have to consider in your country?

The Office of Biological Safety (OBS) in the Department of Environment, Health, and Safety at the UW-Madison runs a biological safety course which was mandatory for all iGEM participants. Beyond adhering to all guidelines established in this, we have not further discussed project specifics with the OBS.

4. Do you have any other ideas 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?

Maintaining best practices established by biosafety committees is always useful, but there are even more interesting options that could be useful for broader synthetic biology safety down the road. For example, making engineered organisms dependent on non-standard nucleotides or amino acids which are only provided in a laboratory setting could help prevent the accidental release of recombinant organisms.