iGEM Safety Questions (Safety Page)

1. Would any of your project ideas raise safety issues in terms of:
• Researcher Safety?
• Public Safety?
• Environmental Safety?

All the bacterial strains we worked with over the course of this project (E.coli K-12 derivatives DH5alpha, DH10b, MG1655Z1, and BW22826) are non-pathogenic to humans, and all proteins produced are non-toxic. Furthermore, proper safety protocols for the handling of these biological materials were always observed, thus minimizing risks to safety and health of the iGEM team members. The general public may include immunocompromosied individuals that may be more susceptible to E. coli K-12, but the risk of pathogenicity remains minimal.

While TALEs are derived from a plant pathogen, Xanthomonas, the TALEs and our chassis do not express pathogenic capabilities. According to the study 'Escherichia coli K-12 Derivatives Final Risk Assessment' published by the United States Environmental Protection Agency in 1997, "the strain E. coli K-12 is a debilitated strain which does not normally colonize the human intestine. It has also been shown to survive poorly in the environment, has a history of safe commercial use, and is not known to have adverse effects on microorganisms or plants."



2. Do any of the new BioBrick parts (or devices) that you made this year raise safety issues? If yes,
• Did you document these issues in the Registry?
• How did you manage to handle the safety issue?
• How could other teams learn from your experience?

The new Biobrick parts that we constructed have minimal potential to generate safety risks. We assembled Biobrick parts that would provide a foundational advance for future iGEM teams and do not pose an intrinsic threat to the environment or to the health of individuals or the public at large. The only concern with the bacteria we used was the small risk that the transformants would genomically acquire the same form antibiotic resistance provided by the plasmids. To mitigate this, we made sure to use sterile conditions when growing these bacteria in cultures and also made sure to safely dispose of any unused cultures. We believe that these precautionary measures are standards by which most labs operate and we hope to remain a model of safety.

It is plausible that our construct could generate biosecurity risks through misuse, but only if extensively reengineered for malicious purposes. Our construct itself does not increase the pathogenicity of E. coli K-12 and does not produce any toxins. Likewise, the implementation of our construct in a large-scale industrial setting should not give way to environmental or biosecurity risks. E. coli K-12 has a history of safe use in a large number of industrial applications. Any risks that may arise will be due to the gene(s) controlled by our system, not our system itself. For example, it may be possible to tailor a pathogen to respond to a metabolite through our system, but this is not a risk intrinsic to our system. Furthermore, the risk for horizontal gene transfer in the environment is remote due to the debilitated nature of our chassis. In any case, our construct comes with multiple points of control, so that there is minimal expression of the gene of interest without the addition of an appropriate inducer.



3. Is there a local biosafety group, committee, or review board at your institution?
• If yes, what does your local biosafety group think about your project?
• If no, which specific biosafety rules or guidelines do you have to consider in your country?

UC Davis has a Safety Services Department on campus that oversees all UCD affiliated research in order to maintain a high standard of safety. We contacted Biosafety Officer M. Malendia Maccree to discuss safety concerns related to our project. She raised no concerns, approves of our project, and has documented it as exempt from NIH Guidelines, which may require initial approval of a project according to Appendices C-II and C-II-A. This exemption is due to the nature of our chassis and the fact that we did not use DNA from Risk Groups 3 or 4.

For more information on our country's biosafety guidelines, please refer to Biosafety in Microbiological and Biomedical Laboratories, 5th edition.


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?

Engineering systems that are orthogonal to the chassis's natural biochemistry could reduce the risk of unintentionally increasing the pathogenicity of the chassis. Systems that are dependent on externally provided metabolites or ligands could also reduce the risk of horizontal gene transfer in the environment. A Biosafety Committee should always be consulted prior to commencing a project that may carry any biosafety risks.