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  • Microfluidics
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• Notebook
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  • May
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• Considerations
  • Human Practices
  • Safety
• Acknowledgements
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Safety

   1. Would the materials used in your project and/or your final product pose:
       

a. Risks to the safety and health of team members or others in the lab?

The aim of our project is to create variants of the TetR transcription factor and to test their binding affinities to E. coli plasmid DNA. Since the nucleotide mutations that are needed to construct these variants are minimal, localized, and well-characterized in literature, there is very little risk of producing dangerous strains of E. coli that might endanger the health and safety of team and lab members. Nevertheless, a great deal of emphasis is placed within the lab on the proper treatment and disposal of bacteria. Both for the experiment's own success and for the safety of others, all bacterial manipulations (cell cultures, glycerol stocks, mini-preps) that involve exposing bacteria to the air of the lab require the use of a Bunsen burner. Moreover, disposing of all bacteria is a two-step process. First, one must bleach the receptacle containing the bacteria (fill its contents with bleach) under the fume hood. The bleached mixture is then poured into a special canister which is treated separately. Second, all cell pellets or dry receptacles containing or having contained cells must be placed in a special biowaste bag, clearly labelled for that specific use. This bag is then disposed of, every week, by a special team from the Life Sciences building. These precautions, coupled with the usual wearing of nitrile and latex gloves at all times, are adequate to the task of maintaining the safety and health of our iGEM team.

b. Risks to the safety and health of the general public if released by design or accident?

The accidental or intentional release of any product derived from our iGEM work is highly unlikely to induce any kind of risk whatsoever. The E. coli strain being manipulated in the lab, the DH5-alpha strain, is entirely innocuous. Furthermore, after any of our manipulations, this strain will only bear resistance to at most one antibiotic at any time (from chloramphenicol, kanamycin, or ampicillin). That being said, hundreds of E. coli strains

c. Risks to environmental quality if released by design or accident?

d. Risks to security through malicious misuse by individuals, groups or states?

       Please explain your responses (whether yes or no) to these questions. 

   Specifically, are any parts or devices in your project associated with (or known to cause): 

       - pathogenicity, infectivity, or toxicity?
       - threats to environmental quality?
       - security concerns? 

   2. If your response to any of the questions above is yes:
       a. Explain how you addressed these issues in project design and while conducting laboratory work.
       b. Describe and document safety, security, health and/or environmental issues as you submit your parts to the Registry. 

   3. Under what biosafety provisions will / do you operate?
       a. Does your institution have its own biosafety rules and if so what are they? Provide a link to them online if possible.
       b. Does your institution have an Institutional Biosafety Committee or equivalent group? If yes, have you discussed your project with them? Describe any concerns or changes that were made based on this review.
       c. Will / did you receive any biosafety and/or lab training before beginning your project? If so, describe this training.
       d. Does your country have national biosafety regulations or guidelines? If so, provide a link to them online if possible. 

   4. OPTIONAL QUESTION: Do you have other ideas on how to deal with safety or security issues that could be useful for future iGEM competitions? How could parts, devices and systems be made even safer through biosafety engineering? 

The 2011 EPFL iGEM team has taken two approaches to its engineering work on transcription factors: one is in vivo and the other is in vitro. Neither presents any form of unusual safety risk. Nevertheless, we should highlight a few aspects of the work that present some risk to the public and to the researcher.

• The in vivo manipulation of E. coli using various antibiotics, like ampicillin and kanamycin, always presents a minimal amount of risk to those who would be exposed to a resistant strain. The highest precautions are taken to manage and control these cell cultures. We dispose of any living matter (cells, pellets, plates) in a yellow bag that is collected every day by the safety units at the EPFL. Prior to disposal, all cell cultures are first bleached.

• The EPFL Safety Committee has put together specific protocols to follow with regards to biological and genetic work that are available on their website. These rules are followed by all EPFL labs including the two labs that organize the iGEM experience.

• As regards chemicals, none of the current protocols require the use of highly toxic materials.

• To get access to the clean rooms where wafers and microfluidic chips (for MITOMI and chemostat experiments) are produced, one has to undergo training with a clean room professional. All iGEM members involved in the production of these chips have been trained accordingly. The website for clean room safety is here. More specifically, the training involves the use and wear of clean room gear (eyewear, body-suits, etc...) as well as the proper disposal of toxic chemicals involved in photolithography.