Would the materials used in your project and/or your final product pose:
a. Risks to the safety and health of team members or others in the lab?
The potential carcinogen ethidium bromide was used in our gel electrophoresis experiments. We followed our labs standard protocols for dealing with the substance, which includes the use of protective material and steps to minimize the use of ethidium bromide. As well, the carcinogen naphthalene was used in our chemotaxis assays. Care was taken to handle all chemicals in a well-ventilated area, using appropriate safety attire.
b. Risks to the safety and health of the general public if released by design or accident?
In any transgenic organism, a potential risk could be posed to human and ecosystem health due to their novel genetics conferring an evolutionary advantage over wild type organisms. Given this, this project is designed to minimize the possibility that such an event could occur. Caenorhabditis elegans is not known to be pathogenic in humans. Furthermore, the extrachromosomal assay used in the microinjection process greatly weakens the constitution of transgenetic C.elegans.
Therefore, if it did escape in the wild, it would be even less likely to be pathogenic to humans than wild type, and would probably be outcompeted by wild type C.elegans in all environments.
c. Risks to environmental quality if released by design or accident?
Transgenic C.elegans would pose a minimal risk to the environment for reasons similar to why they would pose a minimal risk to humans. The extrachomosomal assay previously mentioned makes transgenic C.elegans less robust than wild type worms in most situations. The exception for this would be in an environment heavily contaminated by toxic hydrocarbons. Our transgenic worms would be better at using the hydrocarbons as an energy source. Upon the completion of the bioremediation process however, the advantage possessed by the transgenic worms would disappear and they would once again be out competed by wild type worms. Therefore it is unlikely that the transgenic worms would be able to establish any stable ecological niche. Another concern is that transgenic C. elegans could mate with wild type worms to produce hybrid offspring. While it is possible these genes could propagate through the genetic matrix in this way, they confer an advantage only in the most limited and transient of circumstances. Therefore it is highly unlikely that our constructs would contaminate the C. elegans gene pool with any degree of substance or efficiency.
d. Risks to security through malicious misuse by individuals, groups or states?
C. elegans is a low risk organism for malicious genetic manipulation for a variety of reasons. First, C. elegans is significantly more difficult to work with than a standard E. coli chassis. While E. coli can be engineered with a simple heat shock procedure, C. elegans requires expensive micro-injection equipment and highly trained injectors. Furthermore, C. elegans has never been known to pathogenic to humans, and would be more difficult to modify for such a purpose than a chassis like E. coli. which has well known pathogenic strains. That being said, the novel nature of the C. elegans chassis does carry some risks. The organism has been known to exist symbiotically with some bacteria. C. elegans’ known harmlessness could be a bioterrorism advantage, with the worm acting as a carrier to deliver pathogens past biological detection systems. The worm is also more advanced than E. coli, and is able to access a significant genetic arsenal (via splicing, RNAi, etc.) that is barred from lower organisms. As an eukaryotic organism, the worm is also more robust than its bacterial counterparts. It is insensitive to antibiotics because reproductive nature gives it a greater genetic diversity than most bacteria. A pathogenic C. elegans would be significantly harder to kill than bacteria yielding the same genetic weapons. Regardless, we still think C. elegans is a low risk chassis. While its novel nature does confer some unique options for harmful purposes, a simple chassis like E. coli offers significantly more potential for such purposes . E. coli propagates more quickly, is simpler, and thus more easily manipulated than C. elegans.
Specifically, are there any parts or devices in your project associated with:
a. Pathogenicity, infectivity, or toxicity?
b. Threats to environmental quality?
c. Security concerns?
Our biobricks are considered safe. Many of our parts are naturally occurring genetic sequences in C. elegans and thus cannot be classified as transgenic. Our transgenic components are GPCRs taken from higher eukaryotic organisms such as mice and humans. None of our parts are known to produce toxic or infective effects.
Under what bio-safety provisions will/do you operate:
a. Does your institution have its own biosafety rules and if so what are they?
Queen's University has a very stringent set of bio-safety policies and rules. They are outlined here.
b. Does your institution have an Institutional Biosafety Committee or equivalent group? If yes, have you discussed your project with them? Describe any concerns or changes that were made based on this review.
One one of our Faculty Advisors, Dr. Kenton Ko, chairs the Queen's University Biohazards Committee while Dr. Chin-Sang is the Biology representative.
When discussing the possibility of using the organism Pseudomonas putida in the lab because of it's unique abilities related to biodegradation, were informed by members of the Committee that this organism is a pathogen and a bio-safety level 2. As our labs are only cleared for bio-safety level 1, we took an alternative route and sought enzymes used by this bacteria.
c. Will / did you receive any biosafety and/or lab training before beginning your project?
We undertook WHMIS and Radiation training as part of our commitment to team safety. In addition, no member was allowed into the wet lab until a training session with the lab technician. This training consisted of a variety of different readings on laboratory safety punctuated by tests to ensure we fully grasped the measures involved.
d. Does your country have national biosafety regulations or guidelines?
Canada does have national biosafety regulations. They can be found here.
Do you have any other ideas on how to deal with safety or security issues that could be useful for future iGEM Competitions?
a. How could parts, devices and systems be made even safer through bio-safety engineering?
As a new direction in Human Practices, the Queen’s team is creating a series of recommendations on how the Canadian Security Intelligence Service (CSIS) could deal with bioterrorism informed by synthetic biology.