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Safety in our design

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Safety in our design

For our 2023 iGEM project, we tried to created ARROW, a adipocyte-targeting UCP1-delivering Photorhabdus Virulence Cassette (PVC) to burn out the excess fatty acid in adipocytes. We hope our tool can provide a fancy and safe choice for body-weight control, thereby contributing to the prevention of obesity-related diseases. We successfully confirmed the protein delivery ability of PVC and constructed the UCP1-based fat-burning payload ARROW Head.
Putting safety first, we considered the safety issues as follows:
1) E.coli cells (DH5-alpha and Stbl3) were used for standard molecular cloning. The strains we use have not been reported to cause disease in immunocompetent adult humans consistently, and it does present a minimal potential hazard to laboratory personnel and the environment.
2) All genes cloned into pBR322, pAWP78 and pcDNA3.1 are non-conjugative, preventing the horizontal transfer of our parts. We use P_CMV and Tet as promoter, which guarantees that our bacteria won’t produce large amounts of protein if not induce the expression.
3) All the parts we use are guaranteed to be safe for humans and won’t cause any biological pollution to the environment.

Potential risk

Although the E.coli DH5-alpha, Stbl3, and EPI300 strains we used have not been reported to cause disease in immunocompetent adult humans. However, it presents a minimal potential hazard to laboratory personnel and the environment since most of these cells will be transformed with plasmids with resistant selection markers. If the bacterial wastes are not properly treated, they may spread resistant genes into the environment. So strict limits were set to avoid any strain leakage and human infection.
HEK-293T cells and 3T3-L1 cells cannot survive when they escape the lab. However, accidentally releasing these cells into the bloodstream may cause harmful health issues. So members were careful when modifying cells.
Accidental exposure to or release of toxic chemicals like Oligomycin could generate health or safety hazards to team members or generate environmental hazards. So all the members learned the course Laboratory Safety Protection before entering the lab.
Meanwhile, the cells we use still have the potential to divide. The ethical review of cells with the ability to divide is more stringent. We should restrict the division of engineered cells to guarantee the safety. Another factor is that we should ensure compatibility with other operations like blood draws and surgeries. More lab works need to be done to reduce risks.