Detail the safety and security considerations of your project, adressing potential risks and outlining the measures taken to mitigate them.
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Wet Lab Lead: Christian. Maintaining a safe laboratory environment is essential for protecting researchers and ensuring the accuracy of experimental results. As our project involved sensitive materials such as genetically modified bacteria, plasmid vectors, and chemical reagents, our team carefully evaluated the possible risks associated with each stage of our experimental protocols. Throughout each stage of the project, we followed biosafety guidelines and appropriate procedures to ensure the safety of all team members.
Before participating in wet lab experiments, all team members were required to participate in a mandatory safety training, which comprised two sessions of 3 hours each. The training included both lab safety and biosafety education. During the training, members learned about personal protective equipment (PPE), biological hazards, chemical hazards, emergency procedures, and the proper disposal of biological and chemical waste. This was done to ensure all lab members had an understanding of appropriate lab conduct.
During the training, team members were also familiarized with the specific safety equipment in the laboratory, including fire extinguishers, eyewash stations, spill kits, first-aid kits, and biohazard waste containers. The purpose of this training was to ensure that every team member knew how to respond appropriately if an accident occurred in the lab.
Finally, team members who entered the lab were required to sign a consent form stating that they are informed of the potential risks with our project and have received adequate training to assess and respond to these risks in the lab.
Chemicals
The chemicals used in our project are considered low risk and included common laboratory reagents such as Coomassie Blue. Nevertheless, all chemicals were handled carefully and all team members were informed of proper safety procedures in the case of a chemical spill.
Before using a chemical, team members reviewed its label and relevant Safety Data Sheet. PPE was worn when required. All chemical containers were clearly labeled and kept closed when not in use.
If a chemical spill occurred, team members were instructed to notify a supervisor immediately, prevent others from entering the affected area, and follow the appropriate spill-cleanup procedure. Chemical waste was collected separately from biological and general waste.
Biological Materials
Our project used Escherichia coli strains DH5α and BL21(DE3). DH5α was used for plasmid vector propagation, while BL21(DE3) was used to produce the proteins of interest. Both strains are commonly used laboratory strains and are considered suitable for work under Biosafety Level 1 conditions. A 2000 study by Chart et al. investigated the pathogenic properties of DH5α and BL21 and found that neither did not possess pathogenic mechanisms likely to cause disease. The study concluded that these strains are non-pathogenic and unlikely to survive in human tissues.
Standard molecular biology procedures were followed during all steps of our experiment. All biological materials were stored in closed and clearly labeled containers.
Waste Management
Although the bacterial strains used in our project were non-pathogenic, they were genetically modified and therefore required appropriate containment and disposal.
Liquid biological waste was disinfected before disposal according to the laboratory's established procedures. Solid biological waste, including culture plates, contaminated pipette tips, tubes, and gloves, was collected in designated biohazard containers and sterilized before disposal.
Chemical waste was kept separate from biological waste. Used chemical solutions were placed in properly labeled waste containers rather than being poured directly into the sink.
All experiments were performed within the laboratory, and no engineered bacteria were intentionally released into drains, soil, water, or other parts of the environment.
The laboratory was equipped with necessary safety facilities to reduce the risks associated with experimental work.
All team members were required to follow the laboratory dress code. Laboratory coats, long pants, and closed-toe shoes were worn during wet-lab activities. Long hair was tied back. Gloves were used when handling bacterial cultures and chemical reagents. Team members washed their hands before leaving the laboratory.
Describe all potential safety and security risks associated with your project, including:
Explain how you are addressing these risks and implementing safety measures.
Detail any safety features incorporated into your project design, such as:
Explain the rationale behind these design choices.
Ensure that your project adheres to all safety requirements outlined in the Safety Policies page.
The section below is a separate, independently-written draft, produced by a team member outside the main documentation process. Kept here for reference and comparison; not yet fact-checked by team leadership.
The argument, in full: an engineered organism that processes plastic in a closed cabin only earns the right to exist if the containment story is airtight.
E. coli K-12 (MG1655) is a non-pathogenic biosafety-level-1 organism, a workhorse of molecular biology for roughly 70 years, with well-characterised escape and survival behaviour in the wild — notably auxotrophic for several common metabolites and a poor competitor outside the lab.
Two independent kill switches, both required to be off for cell viability, deliberately built on disjoint failure modes — one transcriptional (thermosensitive λ cI, triggered above 30°C, driving an SOS lethality response), one translational (EL222 light receptor controlling a colicin self-lysis cassette, triggered by 470 nm light) — so their joint failure rate is genuinely multiplicative rather than sharing a common weak point.
| Failure mode | Mitigation |
|---|---|
| Suppressor mutation in switch A | Switch B still active · joint containment OK |
| Suppressor mutation in switch B | Switch A still active · joint containment OK |
| Cartridge breach (mechanical) | Hard cartridge wall + redundant gaskets |
| Horizontal gene transfer | K-12 + low DNA-uptake environment + plasmid-only maintenance |
| Reactor escape during launch | Cartridge sealed pre-flight; switches active pre-flight |
All wet-lab work conducted at BSL-1, with full biosafety committee oversight, minimum PPE of gloves/goggles/lab coat, a spill protocol at every bench, autoclaving of all engineered-organism waste, and weekly safety walkthroughs.
A stated commitment: if a failure mode is identified during the season that can't be mitigated — particularly one suggesting horizontal gene transfer above background — the result would be published, the construct withdrawn from the Registry, and the project ended. Written down in advance, so nobody has to invent that decision under pressure.
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