Contributors
Overview
We believe that it is our responsibility to understand and manage the risks that relate to our activities. Our team ensured that all involved in the wet lab activities understood crucial biosafety rules, such as cartagena laws and release beyond containment, as well as precautions needed when handling genetically modified organisms so that our actions do not lead to any potential harm to others or the environment. Not only that, we made certain that all members, regardless of lab participation, would know about research ethics before our activities began. Overall, it can be said that our team enjoyed these research activities while also keeping within the boundaries of iGEM Safety Rules and Policies.
Chassis Safety
We used E. coli DH5α for plasmid cloning, E. coli C600 for plasmid modification, B. subtilis 168 for conjugational transfer and B. subtilis natto BEST195 as the final chassis. All are classified under Risk Group 1, with B. subtilis especially considered GRAS (Generally Regarded As Safe) and B. subtilis natto regularly eaten for fermentation. Thus, all chassis in this study pose a low risk for humans and the community. During experiments, we followed the biosafety guidelines of our lab according to this biosafety level.
Parts Safety
The aprE gene derives from B. subtilis natto, which is on the iGEM White List. The bacteria is also regularly applied as food, keeping its risks of handling fairly low.
Lab Safety
Before starting any experiments, all members needed to be aware of
biosafety protocols. We watched a video named “Genetically Modified
Organisms Training Materials” that was distributed by Global Science
Campus, a Japanese government-led research organization.
Subsequently, after watching them each member had one-on-one
meetings with our PI to have a two-way interaction to check our
knowledge.
While formulating our project, we all also read the book
“BioBuilder: Synthetic Biology in the Lab” to further deepen our
understanding about not only synthetic biology, but bioethics, its
risks and engineering methods that can be utilized for safety.
As we begin our wet lab activities, we were given a document from
the lab about the safety rules, how to use equipment, and how to
proceed with our research.
In addition, our lab advisors gave us extra rules to follow at the
lab from time to time, thus we members held a group meeting by
ourselves each time to go over the additional safety rules and fully
understand it. The PI and advisors checked each one of us’
understanding and we were not allowed to experiment until we did.
iGEM_Precautions and experimental procedures
Our team also kept a “Basic Lab Protocol” document, which included the error log. We made the basic lab protocol so that our use of lab tools went smoothly and we could minimize errors. Every time we used a new equipment, we made sure to record the process on this document so that others on the team can also be familiar. When we did make errors though, we made sure to write it in the error log so that we can learn from that mistake and not repeat it again. We all made sure to check the document frequently.
Basic Lab Protocol
Japanese ver.
Basic Lab Protocol
English ver.
Basic Lab Protocol_English
In addition, we created a platform on our Discord server where the
wet lab participant of the day reported
・Time
・Experiments
・Reflections
・Next experiment
everyday after wet lab activities. This kept the knowledge
discrepancy between other non-participating members of the day
fairly low, resulting to safer experiments where participants were
aware of which stage of lab activity they were doing.
