"The safety of the people shall be the highest law."
Marcus Tullius Cicero once wrote famously in The Republic and The Laws. And in our lab, we decided that this “highest law” should apply to our experimental procedures as well.
As a project centered around the enhancement of bioluminescence of plants through gene transference, our experiments involved little to no risk to both our experimenters and the ecosystem. Although we are lucky to have a lab with both risk level 1 and level 2 spaces, we are working predominantly in the level 1 space.
Our experiments involved no animal testing, no creation of gene drives, nor pathogens that threaten the human body. This being said, all chemicals, equipment, and machinery involved in our procedures are handled with great care.
All of our members are trained and tested repeatedly before they are allowed to enter the lab. During the training, we are taught with detail the correct method to work with clean benches and sterilized equipment, as well as the step by step procedure of Agrobacterium-mediated gene transfer. We followed strictly the “Laboratories - General requirements for biosafety” issued by General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of China, which is one of the most authoritative sources of laboratory guidelines within the country. To exemplify, we prohibited food in the lab and made sure that all garbage disposed of are autoclaved, so no living bacteria can ever escape the lab.
Professor Xiao, the advisor of our wet lab, has thorough knowledge of our laboratory procedure. With his research focus in synthetic biology, Professor Xiao is extremely experienced with cloning and working with E. Coli. He taught us the procedures and supervised the lab. Dr. Dong, with years of experience with synthetic biology, is responsible for instructing us in day to day experiments. While our assistant professor Dr. Wu, who got his Ph.D. in plant science, guided us in agrobacterium related experiments. Together, they provided double insurance for the safety of our lab.
Project Specific Biosafety
Besides adhering to the general lab procedure, we also have project-specific safety precautions. The lux operon, a cluster of genes that control bioluminescence in the bacterial species Aliivibrio fischeri, we plan to transfer is not proven to be completely harmless to the target plant species like Nicotiana tabacum. So to avoid unwanted gene leakage, we chose Agrobacterium-mediated gene transfer over gene gun and the CRISPR method, completely reducing such risks.
In the cloning and culturing of certain plasmids, we are sometimes working with bacteria with plamid pSB1C3 which has chloramphenicol resistance. If escaped, such bacteria can potentially cause gene leakage to bacteria outside the lab, causing disturbance in the ecosystem. So when we are discarding the waste bacteria with these plasmids, we autoclaved them to ensure that no living bacteria can be released from the lab.
If we introduce the lux operon gene cluster into cells that involved in reproduction, the gene can be inherited, causing unpredictable, unwanted influence in the gene pool of the targeted plant species. So we chose to inject agrobacterium to tabacum leaves instead of establishing stable plants that could stably express our target gene. Therefore, the transformed plants cannot pass the gene and the trait to the next generation.