Our team was aware of the hazards involved with our lab activities, such as e-coli infection and laboratory chemical mishaps, after having discussed and come up with our project. The team was also aware of the environmental safety risks that our genetic alteration of E. coli would pose, so we considered those in our design.
👉🏽 The entire team had to do Labster projects to familiarize everyone with lab work because 77.7% (7 out of 9 members) of our team members had no pre-exposure to biological laboratory experience.
👉🏽 Dr. Elena Rosca, our PI then led the team through a workshop on laboratory safety while she also instructed them on all of the equipment we would be using. The team was prepared to enter the lab following these preliminary actions.
Fig. 1 Laboratory training session with Team's PI
👉🏽 The whole team wore the lab's required personal protective equipment (PPE), such as protective gloves, clothes, lab coats, and safety eyewear, to prevent contamination by chemicals, pathogens, etc.
👉🏽 All workbenches had boxes of gloves and sterilizing alcohol containers on them, and we maintained a lab code that said: "Assume the person who left the workbench before you didn't clean. Assume the person who comes in after you won't clean it."
👉🏽 Dr. Rosca, our PI also ensured that all experiments that required extra caution and bio-safety cabinets were strictly supervised
The team also considered potential accidents that could result in bacteria escaping into the environment, as well as the idea of commercializing the biosensor by:
I. building safety features into our design to ensure the highest level of safety by devising a plan to prevent horizontal gene transfer.
II. ensuring that the bacteria are destroyed if they escape from their confinement. We developed multiple levels of security to ensure the biosensor's containment.
Fig. 2 Diagram depicting levels of security employed for biosensor containment
🦺 Hydrogel Encapsulation
To prevent the biosensor's proliferation and to safeguard it during deployment, we are first encasing it in a hydrogel capsule which consists of Alginate from algae and polyacrylamide. The gel capsule will feature 5–50 nanometer pores when it is manufactured (Tang et al., 2020), enabling the exchange of metal ions between the environment and the matrix solution of the capsule. Because of the nanoscale pores, the E. coli bacteria cannot leave the capsule, blocking horizontal gene transfer. Following the spread of antibiotic-resistant genes to other wild species through gene transfer, environmental hazards could become more serious.
🦺 Methanol Dependency
The second degree of safety is based on including an E. coli strain that is dependent on methanol. The presence of methanol in the capsule matrix is necessary for the methylotrophic E. coli to flourish. Should the capsule unintentionally be broken, and the bacteria escape into the environment, the bacteria will not live since they feed on methanol, which is highly uncommon in the environment
🦺 UV Autolytic Kill Switch
The final security layer focuses on creating a UV promoter that activates genes that produce autolytic enzymes. This makes sure that if any organisms manage to get through the earlier confinement layers, exposure to the UV will assure complete bacterial self-destruction. The UV-induced promoter will start the creation of autolytic enzymes, which will destroy any stray designed organisms.