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

Despite exclusively using non-pathogenic laboratory strains of bacteria, our team endeavoured to further reduce the risk of handling the recombinant microorganisms that we made. This was done by conducting our research in a physical containment 2 (PC-2, equivalent to BSL-2) lab, where all biological material was autoclaved before disposal, to prevent release of these recombinant organisms into the environment. Correct PPE, including gloves and lab coats, were worn at all times, and additional PPE, such as safety glasses, was worn when there was a chemical or particulate hazard. Hand washing was conducted when entering or leaving the lab, and aseptic techniques were adhered to when working with microorganisms.

Additionally, we ensured that none of the genes or parts that we were assembling would act as virulence factors, and that no known pathogens would be involved in our research, with our research plan cleared with the University of Sydney Biosafety Committee. Our team exclusively used non-pathogenic laboratory strains of Escherichia coli for all parts of our project. Although antibiotic resistance genes were used as selectable markers in many of our constructs, it is unlikely that these genes would allow for pathogenicity in the lab strains of E. coli that were used.

2. Do any of the new BioBrick parts (or devices) that you made this year raise safety issues?

None of the parts we created encode virulence factors or toxins known to be a threat to human or animal health, which was confirmed by BLAST searches of each of the sequences. No parts had sequences sourced from known human or animal pathogens. Despite the fact that the integron systems that are seen in nature are commonly associate with antibiotic resistance and virulence genes, these genes are unlikely to be incorporated as gene cassettes into our systems as:

  • Integration of gene cassettes in our system is controllable, and should only occur at significant levels when the controllable promoter is switched on (when arabinose is present).
  • Preferentially, the expression of the cassette would be controllable, with the cassette only being expressed when lactose (or IPTG) is present.
  • All genes were cloned into either pSB1C3 or derivatives of pSB6A1, both of which are non-conjugative, preventing horizontal transfer of our parts.

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

An Institutional Biosafety Committee (IBC) operates at the University of Sydney. All projects involving genetically modified organisms (GMO) must be reported to and approved by the committee. The regulations are derived from the Australian Gene Technology Act 2001. Our project has been approved and complies with the IBC’s biosafety policies and regulations. The School of Molecular Bioscience also has a set of well-documented standard operating procedures.

Our application to the IBC can be viewed here.

With thanks to: