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From 2012.igem.org

Safety Questions

Overall Project Danger Level: Minimal 1. Would any of your project ideas raise safety issues in terms of:

This summer we worked with the common fungi Saccharomyces cerevisiae (yeast) which is already found in a variety of food products such as bread, wine, and beer. S. cerevisiae has a long history of use in food science, biology, and more recently synthetic biology and is known to be non-pathogenic to healthy adult humans. As a result, S. cerevisiae has been designated as biosafety level 1. The CIB1/CRY2 optogenetic system itself is non-deadly in nature and simply produces a fluorescent protein (mCherry) in response to a blue light stimulus. Since Risk = Probability x hazard I’ve provided details about both probability and risk below.

Probability

-Our lab takes lab safety very seriously: from properly separating volatile waste liquids to bleaching all LB broth before disposal. As a result, there’s an extremely low chance of an accidental series of events leading to death, injury, or environmental/property damage.

-The purpose of our project is to create a screening and testing kit for various proteins. As a result, it does not require exposure of our engineered organism to either the environment or human subjects.

Hazard -Our device is in a Biosafety level 1 non-pathogenic organism and serves only to facilitate faster high-throughput protein expression assays. As a result, there’s no clear hazard to human or environmental health.

-Our engineered organism is not infectious, does not produce a toxin, and does not interfere with human physiology or the environment as evidenced by its presence in common foods/drinks such as bread, wine, and beer.

-Mutations in one or several of the parts would simply lead to a broken optogenetic system. Mutations in either the CIB1 or CRY2 proteins would render the dimerization process impossible and a mutation in the Gal4 Promoter would likewise make it difficult for the CIB1-CRY2 complex to induce transcription.

-If our engineered bio-organism was accidentally released into the environment the only potential danger it could have is if it were screening for a dangerous gene or protein that could cause pathogenicity (highly unlikely since the assay is designed to screen for mutations in key human genes).

-Worst case scenario, the organism is released into the environment and is screening for a dangerous protein such as a prion. Prions are an “infectious” protein which can induce other proteins to misfold and become prions. They are linked to several neurodegenerative diseases such as Alzheimer’s.

2. Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues? If yes,

-Parts: Our BioBrick parts only code for either fluorescent proteins or the CIB1/CRY2 fusion proteins whose sole purpose is to dimerize in the presence of blue light. As a result, these proteins are not toxic and are likely only Biosafety Level 1.

-Devices: At the device level our system is no more dangerous than its constituent parts (not very dangerous at all).

-Cell Chassis Enhancement: Absolutely no changes have been made to the cell chassis in this experiment in order to increase it’s survivability.

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

Duke University requires that all projects be passed by a safety committee before being carried out. In addition, the North Carolina School of Science and Math (high schoolers) also have their own safety committee who must also approve of any and all research projects. Before beginning lab work though, each member of the team had to complete a biosafety course (university level) that covered everything from proper lab attire to patient confidentiality and conflicts of interest in case our work overlapped with work being done by the medical center.

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?

I think that proper documentation of parts, devices, etc. in the registry would be the most helpful tool in future iGEM competitions. Proper documentation of parts including safety issues would be extremely helpful and I’ve found that it’s often lacking in the biobrick registry. Earlier reporting of potential biosafety problems and universal standards for biosafety across the country and across the globe would also be helpful.

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