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 genetic materials used in our project pose no safety or health concerns to members in the lab. There are certain chemicals and solvents we use in lab that are known to be toxic but all are treated with proper caution and care.
b. Risks to the safety and health of the general public if released by design or accident?
If released by accident, our parts and materials would do no negative damage to the general public. They are all benign by nature and pose no realistic danger to anyone.
c. Risks to environmental quality if released by design or accident?
The environment is also in no danger from such a hypothetical release for reasons stated above and below.
d. Risks to security through malicious misuse by individuals, groups or states?
No, nothing could be used maliciously because the bacteria we use is in no way capable of causing wide-spread harm or harm to individuals.
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?
Most of the source DNAs used in our project are either from the E. coli MG1655 genome, are commonly used materials in E. coli, or are known genes from RG1 organisms. Though in common use, one of our coding sequences is from RG2 organism: ToxR ToxR and the Pctx promoter are from a virulence island from Vibrio cholerae.1 However, they only encode a transcription factor and a promoter, respectively. Only the N-terminal half of the open reading frame sequence of ToxR was taken from the source sequence, and the Pctx promoter does not include any open reading frames or known coding RNAs. As there is no function to these features except their ability to act as a transcriptional activator and promoter pair, we believe these materials to be RG1. Based on current knowledge of molecular function of these source DNAs there are no specific anticipated risks associated with any combination of these materials beyond the fact that they are derived from RG1 E. coli. The largest single volume used in these studies will be 2 liters, though most experiments involve between 1ul and 40mL of liquid medium.
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.

Although none of our parts require such precaution, we have been sure to take proper care when working with E. Coli. However, none of our parts require special safety documentation beyond general precaution when dealing with bacteria and recombinant DNA.
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.
Yes, UC Berkeley has its own biosafety rules that are regulated by the Environmental & Health Safety (EH&S) office as well as the Committee on Laboratory and Environmental Biosafety (CLEB). Their list of rules are outlined at the link below:
http://ehs.berkeley.edu/healthsafety/biosafety/researchguidelines.pdf
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.
Yes, we have discussed our project with the Committee on Laboratory and Environmental Biosafety, which presides over biosafety in our laboratory. We have filled out their required safety document called the Biological Use Authorization (BUA), which asked us to outline our project and discuss any possible safety concerns our project may present. The document’s questions can be read here: http://ehs.berkeley.edu/healthsafety/biosafety/bua.html#cleb
c. Did you receive any biosafety and/or lab training before beginning your project? If so, describe this training.
Yes, each member of our lab received biosafety training before the project began. This included general laboratory safety, radioactive material user training, biosafety, laser safety and hazardous and chemical spill training. These training sessions are put on by EH&S, and they offer them regularly in the form of a two hour lecture, as well as an online presentation and quiz. Furthermore, each member of the iGEM team was trained by our iGEM Graduate Student instructor on proper lab techniques and safety procedures.
d. Does your country have national biosafety regulations or guidelines? If so, provide a link to them online if possible.
In the US, the CDC works with the National Institutes of Health to publish national biosafety guidelines.
4. 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?
We believe that integrating the safety concerns into new software tools as well as integrating biosafety regulations into general design workflow would improve adherence to biosafety regulations.