We submitted our final iGEM safety form by the deadline on September 7th and it has been approved.
Click here to view our submitted safety spreadsheet.
We are dedicated to following the safety and security rules of our university and the iGEM competition, as well as applicable state and national standards. As part of our project, we also investigated how our project might fit into local regulations which have more strict restrictions on biotechnology than the prevailing national standards. For more information regarding this, please see our Guide to Integrated Human Practices.
We have taken steps to design our experiment in such a way which minimizes risk and exposure to hazardous substances and cells by working in an appropriate setting, and being properly trained by safety officers at our university.
Lab Safety Training and Institutional Requirements
Before work began, all team members were given site specific safety training and a general safety training course for work in a molecular biology lab. Additionally, all Team members have taken in-person Laboratory Safety training for Biosafety, Chemical Safety, Radiation Compliance, Bloodborne Pathogens, Medical Waste, and Laboratory Ergonomics. All team members are certified to use the Biological Safety Cabinet and to work in BSL1 and BSL2 laboratories (although we only required BSL1 to complete this project). Our campus Biological Use Authorization (BUA) committee has reviewed and approved our project involving work with the mammalian cell lines, CHO DG-44 and AML12. We were granted UC Davis BUA #R2565.
The representative environmental toxins that we exposed our bioassay to were 2,4-dichlorophenoxyacetic acid (SDS),warfarin(SDS),and copper sulfate(SDS). Additionally, we used hydrogen peroxide(SDS) as a positive control. Click here for safety measures regarding the spill of hazardous chemicals. Warfarin (GHS code H360) is a reproductive toxin. Click here for the SOP used when handling this chemical. Chemicals were appropriately disposed through the guidelines set by the UC Davis WASTe system. We also collected an environmental sample of water from a tributary of Putah Creek, from Davis, California. This environmental sample was autoclaved and UV sterilized to kill any pathogens present, before working with the sample. Our use of this environmental sample was approved by iGEM headquarters (private correspondence with Piers Millett, Vice President of Safety and Security), and we were told that we were not required to fill out a "Check In Sheet" for it.
Our project involves Biosafety Level One work. The organism and cell lines we are using pose little risk to us, our colleagues, the community or the environment. These risks are further minimized by adhering to the administrative and engineered safety controls in the lab. Administratively, we were all required to take the standard our campus wide safety training, in person site specific training, and additional training pertaining to handling and safe practices with working with BSL-1 organisms and cell lines (see the list of training courses below). Personal protective equipment is used whenever working with these organisms and strains in the lab. All items in contact with E. coli and the cell lines are sterilized by autoclave and/or bleach (final concentration: 20%, 20 minute contact time) prior to disposal. Work with BSL-1 cell lines are restricted to the biosafety cabinet in the lab.
The threats posed by the CHO and AML cells are minimal. Both our cell lines have been categorized as non-infectious and non-toxic. Mammalian cells are optimized to live inside of mammals, and are not likely to survive in the natural environment or infect other mammals.
In order to prevent situations where the chassis organisms escape the lab, all members present in the lab space follow the safety regulations detailed by UC Davis. As an additional precaution all strains are sterilized via bleach (final concentration: 20%, 20 minute contact time) and/or autoclaving before disposal into the general garbage stream.
We used E. coli (non-pathogenic strain DH5α) to clone DNA. E. coli was not permitted to leave the lab and was sterilized via bleach (final concentration: 20%, 20 minute contact time) and/or autoclave prior to disposal.
Our project was intended only to be used in a laboratory setting. We never plan on releasing our transgenic cell lines into the environment. Our anticipated real world application is carried out in a laboratory setting, and the safety protocols that will be followed are the same as the ones we are currently following. We investigated a variety of ethical considerations and how our project might fit into different cultural contexts, and documented these investigations in our Guide to Integrated Human Practices document.
Security and Dual Use Risks
We evaluated our project to have minimal security risks. We did not work with any pathogens or parts originating in pathogens, nor did we produce any biological products which may have security risks associated with them.