Team:Lethbridge/safety

Safety

The University of Lethbridge 2012 iGEM team is using a combined system of cyanobacteria and Escherichia coli to facilitate extraction of unconventional oil deposits while reducing CO2 emissions. We have taken precautions to ensure that our CAB extraction method is safe for the environment, experimenters, and general public.

Researcher safety

The cell chassis being used in this project include various laboratory strains of Escherichia coli as well as the cyanobacteria Synechococcus elongatus PCC 7942. The E. coli strains being used in the laboratory (i.e. DH5α, BL21 (DE3)) are widely used among researchers, non-pathogenic, and designed for use in recombinant DNA experiments. S. elongatus is a non-pathogenic bacterium that has been extensively studied in the research community. To ensure the safety of our team members when handling these organisms, appropriate safety measures are taken including the use of aseptic technique, wearing of personal protective equipment, and properly disposing of waste material at all times. All team members are given site specific safety training for proper handling of equipment and organisms before commencing their work in the laboratory. Controlled access to the laboratory is enforced by keypad entries to restrict access of untrained individuals. These measures will ensure that there are no risks involved in our project when working in the laboratory.

Public safety, or Environmental safety?

Our team has also focused on environmental and public safety through the implementation of our project. As previously mentioned the organisms we are working with are non-pathogenic and are ubiquitous in nature. However, safety concerns are raised when genetically modified organisms (GMOs) are exposed to the environment. We have designed our project so that the genetically modified S. elongatus and E. coli will be optimized for use in bioreactors where their growth can be carefully monitored and controlled while eliminating exposure to the environment. In this way, concerns about contamination of the environment with GMOs are mitigated. As an added precaution, we have designed a series of killswitch modules that can be integrated into the genome of both species to ensure growth only occurs under controlled conditions. If the organisms are released into the environment, the killswitches will be induced to degrade genomic DNA and essential proteins, which will render the bacteria inert and harmless. These measures will greatly reduce any risk of the release of GMOs into the environment, and mitigate threats to the general public.

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?

The BioBricks that the University of Lethbridge 2012 team has proposed to make do not raise any immediate safety issues. However, we have also considered safety issues that may arise due to improper use of these parts. The gene recA, a part of the killswitch module, encodes for a protein involved in homologous recombination. Uncontrolled production of this protein could result in genomic integration of exogenous DNA, which may include antibiotic resistance genes, genes encoding for proteins that produce toxins, or other genetic material that may result in uncontrolled growth of the bacteria. We have chosen to control the expression of RecA through the use of a light-repressible system in conjunction with the killswitch module, thereby only expressing RecA as a means to destroy the bacteria. The use of a scaffolding system to increase the efficiency of an enzymatic pathway, such as acetic acid or glucose production, could raise safety concerns if the targeted pathway generated some kind of toxin or undesirable product. Therefore, these parts should only be used for enzymatic pathways that have been carefully studied, will not produce harmful products, and will not increase the efficiency of side reactions that will produce harmful products. The BioBricks generated for production and export of glucose by S. elongatus do not pose any safety risks, nor do the BioBricks needed for acetic acid production and export. Some of these BioBricks have been used in previous years with no significant safety concerns. There are no direct safety concerns related to our BioBrick parts, but it is important to consider what potential problems could be discovered in the future.

If yes, what does your local biosafety group think about your project?

The Risk and Safety Services department at the University of Lethbridge has an appointed Biosafety Committee, responsible for overseeing and enforcing proper laboratory safety guidelines. Proper training, documentation of laboratory use, and handling of biological and chemical materials are part of their mandate. The committee foresees no problems with the University of Lethbridge 2012 iGEM project, provided we carefully follow their guidelines and procedures. The current Laboratory Safety Manual can be found here:
http://www.uleth.ca/hum/riskandsafetyservices/PDF/Chemical%20Safety%20Manual%20April%2010%202006.pdf

One of the most useful ways to deal with safety issues in future iGEM competitions is the continued use of mandatory project safety reviews. By critically examining the safety issues associated with their project and any new BioBricks, teams can act preemptively to ensure that the researchers, general public, and environment are not put at any risk. Having an appointed safety leader or safety subgroup can ensure that any risks are promptly addressed and dealt with before any harm has occurred. Depending on the particular application of a given iGEM project, teams could approach appropriate industrial, environmental, or government groups to get a second assessment of potential safety risks. These kinds of safety reviews would reflect the opinion of external organizations on the practical applications of synthetic biology and what steps would be necessary to make the project ready for wider use.

To ensure higher levels of safety when working with engineered biological systems, the implementation of control mechanisms can be used. For example, the University of Lethbridge 2012 iGEM team has designed a multi-level killswitch module to be incorporated into the genome of our cell chassis. This will ensure that the bacteria will only grow under controlled conditions and that any contamination of the environment will result in the destruction of the bacterial genetic material and proteins essential for cell growth. By using killswitch modules, uncontrolled bacterial growth in the laboratory or the environment will be prevented.