Team:NYC Wetware&Safety=edit

1. Would any of your project ideas raise safety issues in terms of researcher safety, public safety, or environmental safety?

Our team has taken all of the necessary precautions to ensure the highest levels of safety throughout the project.

The E. coli strain DH5-alpha was used throughout our project. This is a Risk Group 1 laboratory strain of E. coli that have a low, but not nonexistent, virulence. Organisms classified as Risk Group 1 are not know to cause disease in healthy adults. Keeping with registration requirements for Risk Group 1 bacteria, all laboratory work was performed in an environment certified for Biosafety Level 1.

These strains of E. coli are generally not resistant to antibiotics. All bacteria were transformed with bacterial plasmids carrying our desired insert gene and antibiotic resistance to antibiotics commonly used in laboratories for selection (ampicillin, kanamycin, chloramphenicol, etc.), but will not interfere with potential clinical treatment. Neither plasmids nor their insert genes will affect virulence, infectivity or host range. 

Bacteria was cultured in 5mL volumes and all cultures and inoculations were performed in a biological safety cabinet. As part of our project we have coordinated the receipt and library preparation of several experimental Deinococcus strains of bacteria. These strains were cultured by the Daly group at the Uniformed Services University of the Health Sciences, pelleted and sent to us. These strains grow very slowly and at temperatures warmer than the human body, so pathogenicity is not a major concern when working with these species. Regardless, DNA was extracted from them inside a biosafety flow hood with a face pressure of over 100 ft / min. Solid waste (plates, flasks, etc), liquid waste (decanted culture supernatant), and work surfaces were disinfected using 10% bleach before disposal and biohazard disposal bins were used whenever necessary. 

Although we do expect some public concern about our goal of increasing the radioactive resistance of E. coli, we do not anticipate any safety issues with our work. We have no intention of releasing any bacteria into the human body or the environment at any point in the iGEM competition. While some of our speculative future applications would require exposing bacteria conferred with radioactive resistance into the environment, such as for the bioremediation of toxic waste sites, this is of no concern regarding public safety. As stated above, neither our plasmids or insert genes associated with radioresistant pathways will affect the virulence, infectivity or host range. Even if it did "escape" from our lab, the bacteria is effectively harmless. From inception, our team has considered and discussed the possible ramifications of creating bacteria resistant to large doses of radiation. We recognize that the additional resilience of this trait is a potential hazard if transferred to a virulent species. Careful consideration and caution would be necessary before implementing any public application.

2. Do any of the Biobricks or devices we've made raised safety issues.

Our Biobricks to boost the radioresistivity of DH5-alpha E. coli will not affect their virulence, infectivity or host range. For most scenarios, and as long as it only involves non-pathogenic bacteria, increasing the resilience of E. coli when it comes to radiation resistance has no harmful implications. One real issue that we have considered is food irradiation. The process of exposing food to ionizing radiation to kill bacteria is occasionally used by countries around the world. If a pathogenic bacteria were to somehow pick up the ability to resist high doses of radiation exposure, it could no longer be destroyed through means of irradiation. We took the utmost precautions in the laboratory when designing our Biobricks to ensure safety. More importantly, our Biobricks have the potential to solve tremendous environmental issues posed to us today. In the United States alone, the Department of Energy states that there are "millions of gallons of radioactive waste" as well as "thousands of tons of spent nuclear fuel" and also "huge quantities of contaminated soil and water". E. coli hold the potential for breaking down many of the toxic waste to harmless derivates. Engineering E. coli that can function within the radioactive waste would allow for their clean up.

3. Is there a local biosafety group, committee, or review board at our institution?

We have departmental approval for our safety policies, and IRB approval is not needed since we don't have human subjects. As volunteers of Weill Cornell Medical College, we are insured in the case that any accident occurs.