Biosafety

Key Questions

1. Would any of your project ideas raise safety issues in terms of:
   • Researcher Safety:
   • Working in a laboratory environment is not without its risks, so we took a variety of steps to ensure the safety of our team during our lab work. We worked in a laboratory environment certified for Biosafety Level 1 work, with a subset certified for Biosafety Level 2 work. Our work fell within the BSL-1 domain, as the agents used are "not known to consistently cause disease in immunocompetent adult humans, and present minimal potential hazard to laboratory personnel and the environment," as is indicated per [http://www.cdc.gov/biosafety/publications/bmbl5/BMBL.pdf CDC guidlines]. Our protocols and materials were reviewed by the Institutional Biosafety Committee of the Faculty of Arts and Sciences at Harvard, the Committee on Microbiological Safety (COMS), and were found to be in good standing with the NIH Guidelines for Research Involving Recombinant DNA Molecules and the Harvard University COMS policies (see also Key Question #3 below). In addition, all team members were required to attend a certified COMS safety training course for chemical and biological agents.
     Advanced synthetic biology research does at times require interaction with potentially dangerous substances. Listed below are those substances which posed the most significant hazards to researcher safety. Again, however, it should be noted that our use of these substances was consistent with Harvard's stringent safety standards. These substances included:
     • [http://www.chemadvisor.com/harvard/ohsdoc.pl?searchPage=BASIC&search=PARTIAL&entryMode=addMsds&lang=English(US)&mode=base&chem=ethidiumbromide&DB0=checked&file=ohs60703.htm&dbIndex=0 Ethidium Bromide (EtBr)] was used to stain DNA for gel electrophoresis. Although toxic and a suspected mutagen, the harmful effects of ethidium bromide can be avoided by avoiding direct skin contact and inhalation, which can in turn be avoided by proper observance of safety precautions. In order to minimize primary contact, ethidium bromide is directly contacted only with micropipette tips. Skin contact when handling ethidium bromide-stained gels was avoided by the use of nitrile gloves. Secondary contact was avoided by the disposal of ethidium bromide-contaminated gloves and micropipette tips, as well as the designation of a specific bench, fume hood, and set of micropipettes exclusively for use with ethidium bromide. http://www.uos.harvard.edu/ehs/environmental/ethidium_bromide.shtml Additional Safety Information Consulted Regarding Ethidium Bromide
       • [http://tools.invitrogen.com/content/sfs/msds/S33102_MTR-NAIV_EN.pdf SYBR® Safe] is a DNA gel stain which offers a lower-toxicity alternative to its ethidium bromide counterpart. SYBR® Safe was ordered when we began lab work, and was used in place of ethidium bromide as a DNA stain for gels as soon as it became available in our lab in order to avoid unnecessary contact with ethidium bromide.
     • All other toxic substances and chemicals (including [http://www.sciencelab.com/msds.php?msdsId=9927131 chloramphenicol] and [http://www.sciencelab.com/msds.php?msdsId=9925200 tetracycline]) were handled to avoid direct contact, and with observance of proper safety procedures (e.g., Personal Protective Equipment including nitrile gloves was worn at all times within the lab; chemicals and biological agents were disposed only in designated biohazard receptacles).
     • Ultraviolet (UV) light was used in visualizing stained DNA in gel electrophoresis. To avoid exposure to UV radiation when reading gels, protective, UV-blocking shields were used at all times. When performing gel extractions, exposure was avoided by wearing protective clothing to cover any exposed skin and safety glasses with UV-protection lenses.
     • The laboratory strains of E. coli used, all K12 substrains, were non-pathogenic and therefore not a threat to researcher safety. We have conferred various basic antibiotic resistances (including ampicillin, spectinomycin, tetracycline, and kanomycin resistance, all common features of synthetic biology lab work) to our E. coli strains. However, these strains are unlikely to survive in the wild, or in humans specifically—where they would be outcompeted by naturally-occuring bacteria—and thus this resistance does not present a significant problem for researcher safety. In addition, all used materials that contacted the bacteria were decontaminated with 10% bleach, ideally for a 20 minute minimum contact time, before disposal in designated biohazard receptacles.
   • Public safety:
     • We do not anticipate any public safety concerns. As described above, the laboratory E. coli with which we are working are all non-pathogenic, were conferred only basic, controlled antibiotic resistance, and would not be able to survive and flourish outside of the laboratory in any case, as they would be outcompeted by other bacteria. In addition, as mentioned above, all used materials and waste products are disinfected with 10% bleach. Used glassware is also autoclaved to ensure decontamination. Hands are washed upon leaving the laboratory, and a strict one-glove policy is observed outside of the laboratory in order to prevent the public from coming into contact with our chemical and biological agents.
   • Environmental safety:
     • We do not anticipate any environmental safety concerns. Our non-pathogenic laboratory E. coli strains are unable to adequately compete with bacteria outside the laboratory environment. Thus, our bacterial strains do not present an environmental safety concern. In addition, the risk of environmental contamination by the bacteria and chemicals used in our laboratory is prevented by the decontamination and biohazard disposal procedures described above. As mentioned above, hands are washed upon leaving the laboratory, and a strict one-glove policy is observed outside of the laboratory in order to avoid secondary environmental contamination.
2. Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues?
   • No, our created and projected BioBricks do not pose any safety concerns. All E. coli strains in which our BioBricks will be designed expressed are non-pathogenic K12 substrains, and all expressed substances (zinc finger proteins, zinc finger nucleases) are contained within the bacteria and are not secreted.
3. Is there a local biosafety group, committee, or review board at your institution?
   • Yes, our project safety is governed by the Committee on Microbiological Safety (COMS), within the Institutional Biosafety Committee of the Faculty of Arts and Sciences at Harvard.
   • If yes, what does your local biosafety group think about your project?
     • As discussed in Key Question #1, we have presented our project proposal to COMS, and after a review of our materials and procedures, the biosafety office has approved our project and deemed our practices consistent with Harvard and NIH biosafety regulations.
       • To view a copy of our letter of approval from Harvard's biosafety office, please click here
       • For more about biosafety regulations at Harvard, please see Biosafety @ Harvard below.
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?
   • Creating a "Biological Leash": In our project, for instance, we could engineer all our genetically modified bacteria containing rDNA so that they would be dependent upon a specific compound, found only in the lab and not in nature, to survive. In this case, were the bacteria to escape into the wild, they would die as they would not have this compound available to them.
   • A few additional thoughts in response to the general questions posed on the 2011 iGEM Safety Page: Synthetic biology often seems to be perceived in the public eye as a double-edged sword. While advancements in synthetic biology hold many potential benefits, the risks—real or imagined—posed by many synthetic biology-based technologies can, and have, scared people away from investment in or application of such methods. Take for instance, gene therapy. While one school of thought holds that gene therapy is the foundation of a new wave of personalized pharmaceuticals that will revolutionize modern medicine, the same technologies employed in medical gene alterations raise new ethical questions: where do we draw the line between personalized medicine and personalized life? The potential to "customize" the genome, far-fetched or far-off as it may seem, opens another door in the already-complex realm of modern bioethics. While these are difficult questions with no easy answers, we hope that, through the strict observation of the safety practices detailed above, we can play a role in easing the minds of those wary of synthetic biology and continue working to translate the tremendous potential of this fascinating science into tangible, safe, and ethical benefits for all.

Biosafety @ Harvard

Harvard University Committee on Microbiological Safety, FAS Division: http://www.hms.harvard.edu/orsp/coms/Forms/FAS-Forms/FAS-forms.htm
(This division oversees undergraduate research—inlcuding our project—specifically)

Harvard Campus Services, Department of Biosafety: http://www.uos.harvard.edu/ehs/biosafety/
(Harvard's main biosafety page)

Biosafety in the United States

Center for Disease Control and Prevention (CDC)
(The CDC maintains standards for biosaefty and health practices in US industry and research)

• CDC Biosafety Home: http://www.cdc.gov/biosafety/
• CDC Biosafety in Microbiological and Biomedical Laboratories: http://www.cdc.gov/biosafety/publications/bmbl5/BMBL.pdf