Laboratory Safety and Risk Management
We adhered to all iGEM safety requirements throughout the development and execution of our project. Our team conducted all laboratory work at BioCurious, located at 3108 Patrick Henry Drive, Santa Clara, California, in a Biosafety Level 1 laboratory. We performed routine BSL-1 biological work on open benches or in biosafety cabinets when appropriate, and prepared cadaverine and putrescine solutions in a chemical fume hood at a separate location. Before beginning laboratory work, every team member who entered the laboratory completed BioCurious's required safety training and documented their completion through the BioCurious safety training form. The training covered laboratory access and rules, responsible laboratory individuals, differences between biosafety levels, biosafety equipment, good microbial technique, and disinfection and sterilization.
We used several additional biosafety and biosecurity measures. Team members wore appropriate personal protective equipment, including laboratory coats, gloves, and eye protection. BioCurious maintained systems for accident reporting, material inventory, physical access control, and access to laboratory data. There were some potential chemical safety hazards with our project. Cadaverine dihydrochloride presents risks of skin and eye irritation, and putrescine dihydrochloride presents these same risks along with respiratory irritation. We used a chemical fume hood when making stock solutions. For our experiments, we used small quantities of our stock solutions.
Containment and Responsible Technology Use
Our current project is a laboratory proof of concept and was not tested in commercial food packaging. In a future version of the biosensor, engineered cells could be immobilized within a calcium alginate hydrogel positioned beneath the packaged chicken. Liquid released from the chicken would enter the sensing chamber, producing a colorimetric output that could be analyzed using a smartphone. Future development of our project would require a semi-contained release if the engineered E. coli biosensor were tested or used in a sealed chicken-packaging device outside standard laboratory containment, in a commercial setting.
These tests would involve exposing the biosensor itself to chicken exudate while confirming that the engineered cells cannot contact the food, user, or environment. Before any such testing or use, we would need institutional biosafety approval and would need to comply with iGEM release-beyond-containment rules, FDA food-contact packaging regulations, EPA TSCA biotechnology rules, and USDA/FSIS requirements related to poultry packaging and labeling.
We adhered to strict rules regarding AI safety. In the case of our use of Large Language Models (LLMs), we mostly used them to help install software and design experiments. We ran these LLMs in contained environments, preventing them from directly installing software for us in order to mitigate potential risks.
For experimental design, though there is risk of hallucination when using LLMs, we mitigated these risks by only using them to assist in preliminary design. We also paired this usage with stringent team and advisor review to make sure our experimental plans were sound. For generative AI for image and video generation, we only used these images as reference, and all of our actual presented work did not include AI-generated images or videos. We used AlphaFold 3 and Boltz 2, which are standardized AI tools and mitigated risks by fact-checking with other sources.
Expert Consultation and Safety Oversight
During project ideation, we sought help from experts associated with Stanford University who were also potential contacts that we would consult when managing risks. We also actively discussed with the safety committee at our community lab BioCurious, (safety@biocurious.org). Specifically, we had a discussion with the safety committee about handling cadaverine and putrescine before we began our lab work at BioCurious. We used CDC Biosafety in Microbiological and Biomedical Laboratories, the NIH guidelines for Research Involving Recombinant or Synthetic Nucleic Acid Molecules, and Safety at Dow as safety and security rules/guidance.
We have also taken other actions to manage potential risks. Our team conducted multiple rounds of expert outreach throughout the project and incorporated relevant feedback into our experimental and safety plans. We also consulted with stakeholders including food bank workers, meat sellers, restaurants, etc. Lastly, we also modified our experimental design to use the dihydrochloride forms of cadaverine and putrescine to mitigate safety hazards.