Team:Lethbridge/Safety
Biosafety
University of Lethbridge Guidelines
All wet lab experiments are hosted at the University of Lethbridge Chemistry and Biochemistry Department with the exception of home-grown Arthrospira platensis (with the approval of the Safety Committee) which is a food grade product, posing no risk to the public. The laboratory itself is a Containment Level One laboratory which no aspects of our project exceeded these guidelines. Containment level one “requires no special design features beyond those suitable for a well-designed and functional laboratory. Containment is achieved through the use of practices normally employed in a basic microbiology laboratory.”
- Public Health Agency of Canada
Other precautions are taken to further provide a safe work environment for team members. Prior to any lab work students are required to come in lab-appropriate attire, meaning long pants, closed toed shoes and tied-back long hair. Further equipment is given to the students for extra protection such as lab coats, safety glasses, and gloves. All labs at the University of Lethbridge are equipped with fire extinguishers, emergency showers, eyewash stations, and first aid kits in case of emergency situations. WHMIS training is provided to all students before any wet lab experiments, and advisor supervision is provided at all times to further ensure safety. Additionally, a thorough lab safety orientation was given to all students who worked in the lab.
To address biosafety, the University of Lethbridge has a Risk and Safety Services committee tasked with ensuring that a safe work environment is upheld by all labs within the institution. 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.
The Risk and Safety committee is aware of Lethbridge iGEM and of our project. They support our work as long as we follow the Laboratory Safety Guidelines outlined by Health Canada: Public Health Agency of Canada.
Biosecurity - Design Considerations
Light Antenna
We have developed a NAB1 constitutive expression construct (BBa_K3237024) and an antisense NAB1 Dicer-like mediated repressor construct (BBa_K3237012) to manipulate the light antennas of the microalgae. In wildtype microalgae in high light conditions, NAB1 sequesters light antenna mRNA and prevents its translation, truncating the light antenna to prevent photobleaching. By integrating a consitutively expressed, antisense NAB1 repressor system into our C. merolae chassis, we not only enable improved growth efficiency and consistency in a low light lab environment, but would also vastly reduce environmental competitiveness in the wild because the modified microalgal cells would be incapable of modifying their light antennas to adapt to higher light environments and would die from photobleaching.
Kill Switch
To ensure our genetically modified C. merolae strain cannot accidentally contaminate wild populations, we are also planning to implement a kill switch. Celesnik et al. (2016) used expression of the non-specific nuclease, NucA from Anabaena, under a metal-ion inducible promoter to induce conditional cell lethality from DNA degradation. We intend to use a similar system except we will use a zinc-inducible constitutive promoter to induce expression of NuiA, a NucA repressor, to limit NucA expression in microalgae grown in zinc-supplemented media. In the wild, it is unlikely that sufficient zinc will be present for expression of NuiA and repression of NucA will be lifted, enabling DNA degradation and cell death. This system remains to be tested to ensure that sufficient cell lethality is achieved to be an effective biosafety measure.
References
Čelešnik, H., Tanšek, A., Tahirović, A., Vižintin, A,. Mustar, J., Vidmar, V., and Dolinar, M. (2016) Biosafety of biotechnologically important microalgae: intrinsicsuicide switch implementation in cyanobacteriumSynechocystissp.PCC 6803. The Company of Biologists. 5, 519-528.