• Introduction
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  • Results Summary
  • Selection System
  • In-Vitro Characterization
  • In-Vivo Characterization
  • Microfluidics
  • Data
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• Tools
  • Gibson Assembly
  • MITOMI
• Notebook
  • Protocols
  • May
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• Considerations
  • Human Practices
  • Safety
• Acknowledgements
  • Sponsors
  • Partners

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 our 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). While the intentional release of such a strain is an unfortunate possibility, one must recall that hundreds of these harmless E. coli strains are circulating in the average healthy human's intestine at any given time, and that only a handful of existing strains are known to cause severe damage to human health. Since our lab does not use these strains, the risk to the public is minimal.

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

No known environmental damages have ever been attributed to the DH5-alpha strain.

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

As mentioned earlier, there are no risks to security involved in the misuse of these bacteria by any entity.

   2. 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.

[http://sv-safety.epfl.ch/ 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. These are standard rules covering both the ethics and methodology of all biological protocols. Clicking on the hyperlink will redirect you to the appropriate website listing out all of these rules in detail.

       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.

The EPFL does have an institutional safety committee, but not specific biosafety committee. We have been in touch with Stéphane Karlen, the biosciences representative to the institutional safety committee. The modus operandi, as far as project reviews are concerned, is to bring in an expert from the safety committee if toxic chemicals or potentially dangerous organisms are being used by a lab. Since this is not our case, we have not had to present a project review.

      c. Will / did you receive any biosafety and/or lab training before beginning your project? 

If so, describe this training.

We did not receive any specific biosafety training prior to working in the lab. However, whenever a new piece of equipment is introduced or a new protocol is created, we are all briefed on the safety concerns involved in its use. In particular, three of our lab members have received training from the CMI at the EPFL to be able to use the clean rooms for microfluidic chip fabrication

       d. Does your country have national biosafety regulations or guidelines? 

If so, provide a link to them online if possible.

   3. 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? 

• [http://sv-safety.epfl.ch/ 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 [http://cmi.epfl.ch/organisation/security.php 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.