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From 2012.igem.org

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Answers to Safety Questions

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

E. coli strains used: ER2738 expresses F-factor for phage infection with tetR (Biosafety Level 1) DH5a, BL21 (DE3) for plasmid propagation and protein expression (Biosafety Level 1)

S. mutans (Biosafety Level 2) S. salivarius (Biosafety Level 1) S. oralis (Biosafety Level 2) S. mitis (Biosafety Level 2) M13 phage (Biosafety Level 1) Actinoplanes garbadinensis (Biosafety Level 1)

Safety levels retrieved from: www.atcc.org

Our project involves the creation of novel bioactive compounds with the end goal of applying them to the oral cavity of humans. As these compounds have not been tested in humans, their toxicity levels remain unknown and have public safety concerns. The compounds should only be used after extensive clinical trials that are beyond the scope of this project.

Another safety issue brought forth by our project is the use of a targeting peptide for our bioactive compounds. Steps were taken to ensure that the targeting peptide was specific by utilizing negative selection against three oral commensal Streptococci besides Streptococcus mutans. However, as the epitope it targets remains unknown, there is a possibility that the compounds could be targeted to other organisms besides our intended target (S. mutans).

A unique property of our designed bioactive compounds is that they are peptides, and will easily degrade in the environment, unlike several other antibiotics (1). The strains of E. coli used cannot survive outside of laboratory conditions and thus will not allow the engineered genes to enter the environment. 1. Kummerer, K. (2003). Significance of antibiotics in the environment. Journal of Antimicrobial Chemotherapy, 52 (1), 5-7.

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?

Our project involves the creation of novel bioactive molecules through combinatorial biochemistry. This presents a safety issue since there is a possibility of inadvertently producing a molecule toxic to humans or the environment. Although we believe that none of our novel molecules are toxins, it was still important to carefully control their exposure to lab personnel and monitor their disposal. Standard personal protective equipment, such as gloves and lab coats, was sufficient in preventing contact with our uncharacterized compounds and all wastes were disposed as biohazardous waste.

These issues will be documented in full in the BioBrick Registry and other teams should take care to follow our example in maintaining caution when working with novel compounds created through synthetic biology.

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?

At McMaster University, the Presidential Biosafety Advisory Committee reviews and approves projects involving biohazardous materials. Their tasks include, but are not limited to, facilitating legislative compliance amongst McMaster researchers, overseeing the transportation of biohazardous materials and educating both students and faculty on biosafety issues. The Presidential Biosafety Advisory Committee encourages the scientific learning process and supports the projects of the McMaster iGEM team. It is required that students participating in wet lab work adhere to biosafety regulations relevant to the project. This includes a variety of individual safety training before beginning lab work (i.e. biosafety level 2 training). This ensured that students received proper training on safety protocols, the proper use of equipment, and dealing with potentially hazardous waste.

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?

To ensure that iGEM competitions remain safe and are used in the best interests of the scientific community, a database should be created which monitors the potential of all biobricks to become dangerous. There must be a set of regulations that are placed on the use, and even distribution, of these parts to all teams.

Future iGEM competitions can be made safer by restricting modification of pathogenic bacteria and/or genes and requiring safety approval by the iGEM committee upon entry. iGEM teams can be asked to include a section on their website to describe the safety training they were required to complete before working in a laboratory in order to ensure students have acquired proper training, and to emphasize that the science is being conducted in a regulated manner. The safety of parts can be improved by restricting the use of uncharacterized gene cassettes that could potentially be activated by substances readily found in the environment or the human body. In addition, requiring that all bacterial expression vectors contain inducible suicide genes can increase the safety of systems.

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Use this page to answer the questions on the safety page.