Safety Information

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

Our project ideas do not raise any researcher safety issues. One of the dyes that we used is malachite green, which has the ability to cause low concentrations of free radicals as described by J.C. Liao et al . Even though there is a risk of a few free radicals being formed during measurements, all assay-ed micro-plates were properly disposed of and malachite green is relatively safe by itself. We only used non-pathogenic Escherichia. coli, which is a biosafety level 1 organism.. To facilitate the selection of E. coli, we transformed E. coli strains (DH5-alpha and BL21(DE3)) with ampicillin resistant genes. We also used BL21 (DE3) pLysS strain for gene expression, which contains a chloramphenicol resistance gene. While these antibiotic resistant strains may pose a threat to public safety if released from the lab environment, all safety protocols were followed and cells were disposed according to institutional requirements.

Other than the use of antibiotic resistant strains as mentioned above, our project did not incorporate any biological components, which pose a threat to environmental safety. Regarding possible toxic chemicals used such as ethidium bromide for running gels, these chemicals were all disposed of according to institutional requirement. The researchers using ethidium bromide were required to read all of the MSDS forms and participate in chemical lab safety before handling the equipment.

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? issues? If yes,

No, our parts themselves do not pose any risks.

3. Is there a local biosafety group, committee, or review board at your institution?

There is a biological safety group, which is part of the Environmental, Health and Safety department. We have been exempted by this group to perform rDNA work. We obeyed all of the federal, state and local laws pertaining to the disposal of hazardous waste and biohazard waste (including liquids, solids and sharps). Our project used only biosafety level 1 organisms. All of the chemicals we use in the lab have been cleared for laboratory use.

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?

One possible idea is to create an inducible (light-based) “kill-switch” in E. coli and yeast similar to the apoptotic pathways in mammalian cells using LovTAP. This would allow for easy disposal of cells, by simply placing the cells in a dark environment when we do not need them. A similar idea is to create a strain of E. coli with a knocked out metabolic pathway, causing it to depend on a laboratory supplied environment to survive (such as a chemical or light). This would be a more passive safeguard against an accidental release of the bacteria.

J.C Liao, J Roider, D.G Jay. “Chromophore-assisted laser inactivation of proteins is mediated by the photogeneration of free radicals”. Proc. Natl. Acad. Sci. USA, 91 (1994), pp. 2659–2663