Safety questions created by iGEM are designed not only for high school students but all members of the iGEM competition including collegiate and entrepreneurial to remind everyone of the importance of safety even in the competition.

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

For the Lethbridge High School iGEM team we work in a laboratory here at the University of Lethbridge with Containment Level One - which “requires no special design features beyond those suitable for a well-designed and functional laboratory. Containment is achieved through the use of practises normally employed in a basic microbiology laboratory” -Public Health Agency of Canada.

In addition safety in the laboratory setting is a priority concern for us. Before entering a lab setting we are required to wear proper lab attire including a lab coat, glasses and gloves. Long pants, closed toed shoes are required and long hair must be tied back. In every lab we work in at the University of Lethbridge a fire extinguisher, emergency shower, eye wash station, and first aid kit is readily available. We were all informed of where they are located in the labs we work in during our safety training. Although there are some hazardous chemicals we are exposed to or work with such as ethidium bromide, we work under the constant supervision of one of our advisors who assist us whenever there is a potential risk. Before entering in the lab as well we were given WHMIS as well as lab training on the basics of working in a university grade lab.

For our project this year we are working with a non-pathogenic strain of E.coli known as DH5which according to the World Health Organization as rated as a Risk Level One -a microorganism unlikely to cause human or animal disease. In the lab we are mimicking our water treatment system on a small scale by the use of dialysis tubing with pores 1 000 000 daltons big. The pores are large enough to allow the export of just beta-lactamase into the surrounding media but not large enough to allow the E.coli to move across the concentration gradient. If our system was commercialized a similar membrane would be used which would allow just the beta-lactamase to move across the concentration gradient. However we have talked to our local waste water treatment plant on advice for the case where the E.coli is able to move across the membrane. They have advised us that at the end of the process of treating both waste and drinking water, UV light is used to kill any remaining debris and organisms in the water before the use in the public.

Also to address the issue of debris due to cell death, the membrane used in our system would have pores small enough to prevent the debris from escaping to the surrounding media. However we have also designed a safety mechanism if this measure is not sufficient in which we would have a separate pipe leading from our treatment tank composed of a material similar to an EZ-10 column. The filter in EZ-10 columns are positively charged trapping the cell debris, and in our system would apply the same way to manage all cell debris. The debris then would be contained and sent to be treated as biological waste. We have also discussed the safety behind our project with our local waste water treatment plant as well to gain professional and sound advice.

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

Our constructs this year are very similar with the exception of the signal sequences in which we have used Bba_K331007, Bba_K331008 and Bba_K331009. However our genes of interest beta-lactmase and erythromycin do not raise any safety concerns in a lab setting. If all protocols are followed using the BioBrick in the lab the part is safe to use.

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

At the University of Lethbridge to address biosafety Risk and Safety Services is the committee responsible. The goal of the committee is to “... educate, mentor, foster and grow an environment to all members of the University of Lethbridge community where each accepts an individual and shared responsibility in growing a culture that is rich in the assessment, management and control of risk and safety.” -U of L Risk and Safety Services. Their focus is not solely on students but everyone using the campus. For Lethbridge High School iGEM they are aware of our project and support our work provided we follow the Laboratory Safety Guidelines outlined by Health Canada: Public Health Agency of Canada.

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?

Nature.com released a paper which introduces a potential safety issue in the engineering of synthetic bases. Biochemists have developed synthetic bases similar in structure to the four common base pairs of adenine, guanine, cytosine and thymine which have been shown in laboratory results to build DNA that can interact with enzymes. The synthesis of the bases is based on the idea that “ … Rather than hydrogen bonding to keep the two strands of DNA together, these strands nestle up against each other through hydrophobic interactions” -John Timmer. As a result it could be possible to put these synthetic bases into media with growing cells, allow their movement across the cell’s membrane and be incorporated into the cell’s metabolism. For iGEM, the innovation of synthetic bases has potential to be incorporated into project ideas if the theory is successful. Although the innovation may help to reduce safety issues by targeting needed bases for cells, the incorporation of this element poses a risk through malignant use. In addition to the risk of new Biobricks containing synthetic bases being reused by other teams without knowledge to the effects on their systems. To address the issue we propose that if synthetic bases be incorporated into the competition, that knowledge be accessible to all teams and to standardize the practise by a panel of professionals on this field. In addition the creation of synthetic cells does not have a given protocol, and so poses danger in the method and chemicals that could be used. By providing a set protocol on the synthesis of synthetic bases by experts and the biochemists involved it will allow others to work safely with given instructions.