Contents 1 Safety 2 Safety Questions 2.1 How we address safety issues in terms of: 2.1.1 Researcher Safety 2.1.2 Public and Environmental Safety 2.2 Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues? 2.3 Local Biosafety Regulations 2.4 Canadian Biosafety Regulations 2.5 Ideas for how to deal with safety issues that could be useful for future iGEM competitions? 2.6 How could parts, devices and systems be made even safer through biosafety engineering?

Safety

Our University of Lethbridge iGEM 2010 team is actively involved in developing a synthetic biology-based approach to bioremediation of the tailings ponds. In line with this, we have developed guidelines, which dictate what can and cannot be done in the laboratory. This is all done to ensure the safety of the experimenters (students), the environment, and the public as a whole.

Safety Questions

How we address safety issues in terms of:

Researcher Safety

As the Lethbridge iGEM team, we primarily use the BL21 (DE3), and DH5α, strains of Escherichia coli bacteria. Being non-pathogenic, these bacterial strains are the most widely used among biologists, biochemists, and biotechnologists. Possessing unique qualities that make them ideal for DNA recombinant experiments, BL21 (DE3), and DH5α E. coli cells are regularly employed specifically, for protein overexpression, and transformations. These experiments are the foundation for the majority of the projects in the iGEM competition, and are one of the main reasons why these particular E. coli strains are used preferentially by a large proportion of laboratories globally.

Many other precautions are also taken in our laboratory. Aseptic technique is maintained at all times and experimenters wear the appropriate protective laboratory clothing such as lab coats and safety glasses. As well, experimenters dispose of any harmful waste chemicals, such as chloroform, in the organic waste bucket located in the fume hood. Any liquids or solvents that come into contact with bacteria, are disposed of in a large vessel containing bleach. These measures ensure that there are no risks associated with the implementation of our project in the lab.

Public and Environmental Safety

The safety measures employed in our laboratory are designed to cater not only to the needs of the experimenters, but are also intended to consider the concerns of the general public. Introducing an engineered E. coli that will degrade harmful chemicals in tailings ponds naturally raises concerns for public and environmental safety. However, as was previously discussed, the specific strains of E. coli used in our project are not harmful to humans and other organisms in the environment. Additionally, they form part of the Escherichia genus, which is ubiquitous in nature. Furthermore, the team plans to incorporate the gene BamHI into the bacterial genome, which will enable degradation of the bacteria’s genome once it has been released. In the future, the team aspires to localize the catechol degrading enzyme catechol-2,3-dioxygenase into microcompartments , which can then be distributed in the form of a biodegradable powder. This will eliminate the use of bacteria altogether and should greatly alleviate the concerns of public and environmental safety.

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

The BioBrick components that the 2010 Lethbridge iGEM team has made do not raise any immediate safety issues. However, as a team, we have examined possible future consequences that could arise from the improper use of any portion of our submitted parts. The Mms6 gene could be used to generate toxic magnetic nanoparticles that if found in a high enough concentration could potentially pose a risk, especially if ingested. Although the gene, xylE, is not particularly harmful on its own, the chemical compound, catechol, that the xylE protein catechol-2,3-dioxygenase is responsible for breaking down, can be poisonous upon ingestion and therefore appropriate safety measures should be taken. Finally, the microcompartments made from the lumazine synthase gene, could serve as potential storage vesicles for inappropriate agents. Even though no safety issues related directly to our BioBrick parts were evident this year, it is important to consider what future teams or individuals may discover.

Local Biosafety Regulations

At the University of Lethbridge, the Risk and Safety Services department has appointed a committee devoted to biosafety. This university committee ensures that biological materials are used safely on campus, and foresee no problems with the Lethbridge 2010 iGEM team’s project, as long as the proper safety practices in the laboratory are employed.

Canadian Biosafety Regulations

In Canada, biotechnology being released into the environment to clean up the tailings ponds, is regulated by both Health Canada and Environment Canada who share the responsibility for the risk assessment under the Canadian Environmental Protection Act. This includes the risks to human health and safety and environmental impact of the biotechnology. Assessment takes place prior to full scale manufacturing of the product, and if a risk is found, measures are taken to reduce that risk by either banning the product, or introducing regulatory procedures.

Ideas for how to deal with safety issues that could be useful for future iGEM competitions?

A way to address safety issues in future iGEM competitions could be to add a mandatory safety review for each part being submitted to the Registry of Standard Parts. This safety review would include all potential risks, uses and any other relevant information for the part that could pertain to its potential safety hazards.

Additionally, one member of the team could be in charge of ensuring that the entire team is aware of any safety issues that could potentially be associated with their project. In order to do so, the designated safety person would need to gather the necessary reactions and thoughts from both their fellow team members and the public to determine how synthetic biology and their project in particular could pose a danger.

How could parts, devices and systems be made even safer through biosafety engineering?

As previously mentioned, the 2010 Lethbridge iGEM team has proposed the incorporation of the BamHI gene into its constructs, which would allow for degradation of the bacterial genomic material, upon release into the tailings ponds environment. In general, in order to make biological engineering safer, having control over the growth of the system could be accomplished through the careful planning and design of the parts and devices that comprise them. This could be established by having bacteria incorporate a plasmid that can be triggered to translate a toxin. By choosing these toxins to be endonucleases, scientists can destroy the genetic material within their bacteria and therefore prevent future replication.