Team:Tokyo-NoKoGen/Safety

What is the risk of our project?

　　In our project, we expect the cockroaches to eat the trehalase inhibitor, 3,3’-dkT, synthesized by our engineered E. coli on a solid medium. However, one possible problem is that E. coli on the surface of the roach’s body or legs may be carried outside of the trap and diffused. The E. coli may potentially spread in the environment and affect the ecosystem by acting on other insects.

　　To help prevent the unwanted spread of our engineered E. coli, we expect the future addition of a light-inducible lysis device. This would allow the E coli to grow well and produce the insecticide while in the trap in the dark. However, any E. coli that exits the dark roach habitat would die once exposed to light. A possible arrangement may involve placing a T4 phage lysis cassette (T4 endolysin and T4 holin) under the control of a light-inducible promoter system (e.g. green-light inducible CcaS/CcaR two-component system).

Our training

We have already had safety training.

All members are given a detailed training about basic lab safety, such as how to safely handle reagents and equipment (autoclave, burners, etc.). Every week, we have a short meeting to discuss any safety issues that come up.We also learn how to handle genetically modified organisms during experiments and how to treat before disposing them.

>Laboratory safety training requirements of our institution (Japanese only)

Our local rules and regulations

Dr. Akinori Koukitu is responsible for biological safety at our institution.

We submitted the details of our projects to him. We follow his advice.

>Biosafety guidelines of our institution (Japanese only)

>The regulations that govern biosafety in research laboratories.(Japanese only)

Risks of our project

The safety and health of team members or other people working in the lab will not be affected by our project.

The only species we use is Escherichia coli, all of which are classified as risk group1 and they do not harm human health.

In our project, our engineered E. coli will produce 3,3-diketotrehalose, which is an inhibitor of trehalase.

However, this compound will not affect our health because we do not use trehalose as an energy storage. General public will not be affected by our product. Even if our E. coli is ingested by mammals, it should not cause any harm because they do not use trehalose as an energy storage.

The environment will not be affected by our product. If the biological materials escape from our lab, some insects that live on trehalose might be affected by the enzyme inhibitor.

However, E.coli that we handle will not survive long outside the laboratory because our E.coli will have a lysis device that responds to light, causing the E. coli to die if it escapes from our lab. Our engineered E. coli is not expected to have any harmful effects on mammals, so there should not be any security risks.

Furthermore, because the E. coli will have a light-induced lysis device, the cells are expected to rapidly die if maliciously spread into the environment.

To minimize risks to team members, we have a strict policy that anyone who enters the lab must wear goggles and lab coat, and anyone doing experiments must also wear gloves. Eating and drinking is strictly forbidding in our lab.

To minimize risks to public health and environment, we ensure that all biological materials are promptly autoclaved after use and disposed of appropriately. Furthermore, we follow our University and country regulations and guidelines for collecting and disposing of any toxic waste generated; our facility is already equipped to handle such wastes.

Biological materials and reagents used in our experiments are never brought outside the laboratory.

To minimize the risks of malicious misuse, the lab facilities are only unlocked when experiments are carried out, so our Biobrick parts and reagents cannot be stolen.