Would any of your project ideas raise safety issues in terms of researcher safety, public safety, or environmental safety?Our team has worked with painstaking care to ensure the highest levels of safety throughout our project.
The E. coli strain DH5-alpha was used throughout our project. This is a Risk Group 1 laboratory strain of E. coli that has a low, but not nonexistent, virulence. Organisms classified as Risk Group 1 are not known 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. Nevertheless, DNA was extracted from these samples 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. Even if it did "escape" from our lab, the bacteria is effectively harmless; as stated above, neither our plasmids nor our insert genes will affect the virulence, infectivity, or host range. 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.
Do any of the Biobricks or devices we've made raised safety issues?One serious issue that we have considered is food irradiation. The process of exposing food to ionizing radiation to kill bacteria is used in countries around the world. If a pathogenic bacteria were to acquire the ability to resist high doses of radiation - a trait we hope to Biobrick - it could no longer be destroyed through this means of irradiation.
With deference to this safety concern, we chose to pursue our project because of its potential for ameliorating a major environmental crisis. In the United States, 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". The biobricks we hope to design would enable E. coli to survive while breaking down nuclear toxic waste to harmless derivates.
Additionally, the ability to increase cellular radioresistance has beneficial medical applications. Fortifying stem cells against radiation damage could increase the success rate of stem cell transplants, a current therapy for patient suffering from diseases of the blood, bone marrow, and certain cancers. We believe that the urgency of such needs justifies our research, notwithstanding its potential hazards.
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.