Lab Safety

For the 2025 Toronto iGEM project, our team ensured that all lab work was undertaken in organized lab spaces and with proper safety training. Our team worked across three different lab spaces this year (two BSL1 spaces and one BSL2 space), requiring proper communication and adherence to best safety practices.

To work in the lab, our team completed safety training courses from the University of Toronto’s Office of Environmental Health and Safety (EHS), as well as departmental-specific safety training from the Chemical Engineering (ChemE) department to work in BioZone (one of our lab spaces). These trainings provided a solid foundation on general biosafety practices, WHIMS practices, wearing PPE, and dealing with accidents and incidents within the lab.

Further, members doing wet lab work needed to receive lab tours and equipment-specific training. The details of these processes are elaborated below.

Safety in BioZone

For work in BioZone, one of our lab spaces, all team members began by completing the required EHS and ChemE safety modules and submitting Research Registration Forms (RRFs) and the Memoranda of Understanding and Agreement on Biosafety (MOUs). A 6-hour online module specifying all operational risks, safety, emergency situations, and proper lab attire was completed prior to in person training. Then, each member participated in safety interviews with our team’s primary PI, Dr. Christopher Lawson. During these interviews, Dr. Lawson reviewed our documentation and assessed our understanding of departmental protocols, institutional policies, and federal biosafety regulations, including lab-specific hazards and emergency procedures.

After successfully passing this assessment, we attended an in-person safety orientation led by Endang (Susie) Susilawati, a ChemE Safety Committee member and BioZone lab manager. This session included guided tours of the BioZone facilities, where we were introduced to the physical layout and trained in the proper use of emergency equipment such as eyewash stations, safety showers, spill kits, fire exits, and first aid kits. We also received hands-on instruction in the safe operation of essential lab equipment including autoclaves, centrifuges, and fume hoods, and reviewed protocols for waste disposal, reagent storage, and workspace maintenance. Throughout the orientation, all team members wore appropriate Personal Protective Equipment (PPE) – lab coats, gloves, goggles, and closed-toe footwear – and practiced aseptic technique and clean lab maintenance. Only after completing all training steps and demonstrating full compliance were we granted permission to work independently in the BioZone laboratories.

All team members understood project-specific risks and their voluntary participation in research activities. We reviewed all applicable Safety Data Sheets (SDS) and Pathogen Safety Data Sheets (PSDS), and committed to wearing appropriate Personal Protective Equipment (PPE) at all times, including lab coats, gloves, goggles, and closed-toe footwear. We reported any incidents, exposures, or safety violations to our primary PI, and ensured that we cooperated fully in any investigations. Through this process, we demonstrated our commitment to maintaining a safe and responsible research environment.

Safety in the BME Teaching Lab

For work in the Biomedical Engineering (BME) Teaching Lab (another of our lab spaces), members followed a similar process as above, completing EHS safety modules, taking an in-person tour of the lab space prior to any work, and ensuring that best biosafety practices were adhered to.

Safety in the MaRS lab space

Work in the MaRS lab space (located in the MaRS Discovery Centre) was primarily undertaken by graduate students on the iGEM Toronto team currently part of the Maxwell and Davidson labs. These graduate students have years of experience working in the MaRS lab space, as well as deep knowledge of proper biosafety procedures and practices. Other members of the iGEM Toronto team doing wet lab work in the MaRS space were supervised by these graduate student members of the team to ensure adherence to proper biosafety.

White List requirements

To ensure compliance with iGEM’s White List, our team submitted HK97, P2 and Mu bacteriophages for approval via iGEM’s Check-In form and got confirmation. All other organisms, biological parts, and proteins utilized in our project are on iGEM’s official White List.

Safety practices during lab work

All our experiments were performed in a biosafety level 1 or 2 facility, with appropriate Personal Protective Equipment (PPE). All our laboratory facilities are organized and spaces are thoroughly documented with proper labelling.

All team members maintained aseptic technique to prevent contamination. Work was conducted under a flame to create a sterile field. In the BME Teaching Lab, we worked under an ethanol flame as this facility did not have a natural gas line. In BioZone, we utilized a bunsen burner. An extra gas valve is in place at the back of the room for safety. When necessary, the team worked inside a biosafety cabinet.

All flammables and corrosive chemicals are stored in chemical cabinets in the chemical room in BioZone. Any chemicals that were aliquoted from their original container were labelled with the WHMIS identifiers to ensure safe handling.

Solid and liquid waste were discarded into designated containers. Biowaste, chemical waste, contaminated glassware were all discarded separately into different containers for proper disposal. These waste bins were emptied when they were ⅔ full to prevent spillage.

All our wet lab team members were trained to operate BioZone’s autoclaves. Solid waste was discarded into the designated containers then autoclaved. Liquid biological waste was discarded into our waste bins based on the risk of chloroform formation. Non chloroform risk liquid waste was bleached and disposed of, whereas chloroform risk waste was discarded.

Phage work was done solely in the MaRS building, as the Maxwell and Davidson labs have dedicated facilities and expertise for handling them. This prevented any cross-contamination between

Anticipating and mitigating future risk

In the spirit of promoting safety with our project, we anticipate future wet lab risks that could be associated with our work, as well as how these can be mitigated.

Chemical/biological hazards and accidental exposure

In the future, we plan to test many promising RBD candidates generated by the dry lab team’s model. Working on assays like Gibson Assembly and RBD swapping to clone and validate these sequences increases risk of exposure to chemical and biological materials.

To mitigate this risk, we will ensure that our members are even more thoroughly trained in cloning and phage work to prevent any risk of personal or cross-contamination. This involves adhering to proper standards of aseptic technique, PPE usage, accident/incident reporting, and communication with lab personnel.

Ergonomic hazards

Frequent lab work poses ergonomic hazards. To avoid these, wet lab members are encouraged to learn about and maintain proper posture during lab work, and to avoid manually doing experiments with extremely large numbers of samples. One strategy is to distribute work amongst multiple lab members simultaneously for a given experiment to reduce ergonomic strain.

Accidental transmission

Working with lysogenic phages (as we had done in this project) poses key risks. In particular, these phages can integrate themselves into bacterial genomes, spreading amongst bacterial populations. In clinical settings, this makes it possible for these phages to propagate in the absence of extremely tight containment procedures.

Thus, once moving past our proof of concept stage, we will switch to using only obligately lytic phages, as these have a significantly reduced risk of genome integration and uncontrollable spreading. As such, we would explore different recombination systems for these lytic phages (such as with yeast) instead of our current RBD swapping procedure (the latter which relies on our candidate phage’s lysogenic nature to permit recombination).

Misuse

Using PHORAGER for unintended or nefarious purposes (such as creating phage RBDs to target healthy bacteria) is a future concern. To mitigate this, all members authorized to use this model should be well versed in phage biology and ecology to understand the consequences of off-target phages being released into the environment.

Having strict controls on who is authorized to use the phage bank, what phages are present in the bank (such as only lytic phages, phages with low transduction potential, etc.) and multiple in silico validation steps to ensure the safety of the generated proteins will be crucial. We are also planning future checks in the wet lab that involve automated large library screening of generated phage candidates. This involves assessing the binding and infectivity of mutant phages against large collections of bacteria, which should provide an in vitro assessment of the phages’ host ranges. This further enables using only phages that are safe.