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

Throughout the project, we worked hard to ensure high levels of safety. The organisms we used in our project include E. coli strains DH5-alpha and BL21 DE3 and Synechocystis sp. PCC 6803. These organisms are classified as Risk Group 1 laboratory organisms and are unlikely to cause pathogenic harm to the team and members of the lab.

We derived our genes of interest from Synechocystis sp. PCC 6803 through colony PCR. Because the organism itself was not modified and is a common freshwater cyanobacteria, it is not expected to raise any risks.

Importantly, E. coli (DH5-alpha) are not expected to survive in the environments other than under specific laboratory conditions, and thus if containment issues fail and modified bacteria are released, they would not be expected to cause an environmental hazard.

Our team followed basic laboratory procedures appropriate for the BSL-2 laboratory we were working in. Due to the nature of this project, some basic safety features have to be taken into account when in the lab. Always, in the lab, gloves, lab coats and safety spectacles were worn to prevent any problems through cross contamination and removal of vectors or bacteria from the lab to the public environment.

When using pipettes in the lab, all pipette tips were discarded into a central beaker and then discarded into a biological waste bin. Any pipettes that were used to transfer bacteria were transferred to a beaker containing 5% DECON to kill the cells before disposal of the tips into the biological waste bin. This process was used for any cell containing vessels; falcon tube and glass vials were washed before being disposed of or autoclaved.

After running DNA gels the gel itself was thrown out into a chemical waste bin for disposal, and the TAE (Tris base, acetic acid and EDTA) buffer was poured into a large container. The reason for this is because they both contain ethidium bromide (EtBr), due to toxicity, carcinogenicity, mutagenicity, and skin irritant. The ethidium bromide is then collected and disposed off in the chemical waste bin, whilst the buffer was poured down the sink.

Several of the protocols followed included potential risks because of the use of hazardous substances (Please refer to: Lab book ). Team members signed a Hazardous Substances Risk Assessment Form, before they were able to begin any of their lab work, see below for more information.

We do not consider our project could raise any risks to security from malicious misuse.

To conclude, we believe our project does not pose any risks to team members, publics and environment as the members of the team have been trained in lab safety protocols.

2. Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues? If yes, did you document these issues in the Registry? How did you manage to handle the safety issue? How could other teams learn from your experience?

None of the new BioBricks we’ve created raise any safety issues.

3. Is there a local biosafety group, committee or review board at your institution? If yes, what does your local biosafety group think about your project? If no, which specific biosafety rules or guidelines do you have to consider in your country?

Our supervisor falls under the requirements of the University Chemical and Biological Hazards group which fulfils both the requirements of the University Court and the Principal’s Office as well as the legal requirements pertaining to biological agents and GMOs (genetically modified organisms).

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?

We believe there should be an iGEM-specific online discussion group, where teams could post their concerns about potential biosafety issues that their (or even some other team’s) research may cause. Through discussion with other teams' students and advisors, a consensus could be reached on whether or not the project is safe to pursue. Additionally, risky sequences could be detected if a screening system is developed and available to use by all iGEM teams as a quick way of ensuring biosafety.

Hazardous Substances Risk Assessment Summary Forms