As responsible researchers, we must choose suitable organisms for experiments, refrain from using organisms in risk groups 3 and 4, adhere to the non-release policy, perform experiments in appropriate laboratories, and ensure personal protection during experiments. Here is WIST’s safety information:

Biosecurity

toxin antitoxin kill switch.

to avoid any artificial pollution to the environment, we designed a killswitch that can be operated by us to kill Bacillus subtilis after it completes its job. This killswitch system is inspired by previous iGEM team, iGEM17_CCU_Taiwan. We made improvements and modifications so that it is more compatible with our project.

MazF induces cell death by precisely cleaving mRNA at a specific location, whereas mazE serves as an antitoxin, countering the effects of the toxic protein MazF.

the killswitch is only activated when lactose is applied.

To exert additional control over this system, we've integrated a lactose inducer based on the lac operon, controlled by the constitutive promotor: pveg.

This operon allows the translation of the coding sequences only when lactose is present. In situations where lactose is lacking, both genes' translation is halted, resulting in the faster degradation of mazE compared to mazF. As a result, mazF is left unrestricted, ultimately leading to cell death. This design not only ensures the effective functioning of the kill switch but also enables activation within controlled timeframes.

this diagram illustrates the condition where the killswitch is not activated

the protein produce by LacI will block the way of mazE, mazF, therefore repressing the expression of the killswitch

this diagram illustrates the condition where lactose is presented. The lactose will bind with the protein LacI produced, then leave the
operator. The killswitch is then activated.

Chemical Control

In our experiments, we use and dispose of various chemicals in accordance with NCHU’s Safety Center regulations and guidelines (officially known as the Center for Environmental Protection & Occupational Safety and Health). All experimental procedures are conducted within a fume hood, and personal safety protection is provided (refer to the "Protective equipment" section for details).

Protective Equipment

Using appropriate personal safety protection equipment (PPE) is essential during experiments in qualified environments. We utilize gloves, goggles, and lab coats for added safety.

Disposal

Biological waste is sterilized in an autoclave at 121°C for 20 minutes and disposed of in accordance with NCHU Safety Center rules. Chemical waste is handled by university officials who transfer it to a licensed third party for disposal.

Safety Training

All wet lab members at WIST undergo comprehensive laboratory training, which covers topics such as pipetting techniques, safe handling of microorganisms, emergency equipment usage, and proper procedures for high-speed centrifugation.