"

From 2011.igem.org

Safety

1. Would the materials used in your project and/or your final product pose:
   • Risks to the safety and health of team members or others in the lab?

   While our project is not outstanding in the amount of danger it poses, there are risks in standard practices that we as researchers need to be aware of.

   In terms of the chemicals and techniques used in our lab, we regularly handle ethidium bromide (EB) agarose gels for imaging DNA fragments. To avoid exposure to EB, there is a designated bench area and set of instruments that are only used for processes involving EB. When working in this area or using equipment that has come into contact with EB, exposure is avoided through the use of nitrile gloves. Similarly, exposure to UV light is avoided by using appripriate shielding from UV lamps. When doing gel extraction, a face shield and appropriate clothing are worn to prevent exposure.

   We also handle polyacrylamide gels for imaging protein. These are handled in a similar way to agarose gels.

   The workhorse organism of our lab is E. coli Top10, a multideficient strain of E. coli that does not thrive outside of a lab environment. The next most commonly used strain in our lab is the environmental bacteria Caulobacter crescentus, a species of bacteria that poses no threat to humans as it cannot survive at either the salinity of the human body nor the temperature. Additionally, even though Caulobacter is a gram negative bacteria, it produces so little endotoxin that it does not produce a noticeable immune response. More dangerous are Staphylococcus aureus and Staphylococcus epidermidis, though both of these are commensal on human skin, in human nasal cavity, and in the mouth. Strains of E. coli Top10 and Caulobacter are given antibiotic resistences for selection purposes, but this is within normal lab protocols. Outside of this, the strains we are using are not given any extra ability to grow outside of the lab environment.

   We take care to autoclave all waste that comes into contact with biological materials and sterilize our benches with ethanol. All glassware is bleach sterilized before being washed.

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

   The strains we are using are non-virulent, and the constructs we have engineered do no increase the fitness of our strains outside of the lab environment.

   • Risks to environmental quality if released by design or accident?

   Our working strain of E. coli does not thrive outside of a lab environment due to being multideficient. Caulobacter is a bacteria native to the environment, and as such care is taken to sterilize so as to avoid the accidental release of the lab strain into the environment. The strains of S. aureus and S. epidermidis are both unchanged from the wild type.

   Genetic material and proteins are also autoclaved before they are thrown away. Small amounts of EB are regularly used, so there is a special waste protocol for used agarose gels.

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

   We work in a locked lab which people outside of our lab group and immediate support staff lack access to.

   Malicious use of the parts we have created in and of themselves and in the combinations that we have engineered pose no threat to the security of individuals, groups, or states.

   • 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?

     The parts we have designed do not cause any safety concerns.

2. If your response to any of the questions above is yes:
   • Explain how you addressed these issues in project design and while conducting laboratory work.

   Answered above

   • 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?

   The current editions of the National Institutes of Health Guidelines For Research Involving Recombinant DNA Molecules and the CDC/NIH Biosafety in Microbiological and Biomedical Laboratories.

   • Does your institution have its own biosafety rules and if so what are they?

   An institutional Biosafety Committee (IBC) comprised of the College faculty and staff and two outside community members fulfill the responsibilities described in the National Institutes of Health Guidelines for Research Involving Recombinant DNA Molecules.

   • 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.

   No changes were made based on this review.

   • Will / did you receive any biosafety and/or lab training before beginning your project? If so, describe this training.

   Yes, we were instructed how to protect ourselves from potential dangers in the lab such as ethidium bromide, UV light, sharps, and what to do in case of a biological spill.

   • Does your country have national biosafety regulations or guidelines? If so, provide a link to them online if possible.
   There is oversight in our country:
   • The Office of Biotechnology Activities at the NIH
   • And theCenters for Disease Control and Prevention