Prologue

As responsible researchers, it is necessary for us to select appropriate organisms for experimentation (which includes avoiding the use of organisms in risk groups 3 and 4), comply with the non-release policy, conduct experiments in appropriate laboratories, and take proper personal protection in the experiments. The followings are NCHU_Taichung’s safety-related information:

Biosecurity

1.Biosynthesis of PQQ:Within our plasmid, we have inserted a blue-light-induced kill switch, comprising a light receptor protein, a toxin-antitoxin (TA) system, and a LacO/LacI regulation system. Our choice of these parts is strategic, primarily aimed at preventing environmental release. The functionality is triggered when the light receptor protein responds to blue light, subsequently inducing the release of toxins to eliminate the host organism.
2.experiments on bee:
We have established safety measures throughout our experiments to prevent any potential problems or accident. For examples: all assays are conducted within separate containment hoods and cages. Moreover, we manage the risk by conducting skin tests for wetlab members before bee experiments. By injecting a small amount of bee venom on the skin, we can observe whether it would cause allergy to the member, without having a severe reaction. Additionally, wearing protective gear, such as anti-sting gloves, is another method we take to manage the risks.

Chemical Control

Among the chemicals used in the experiments, including acetic acid, sodium hydroxide, phenol: chloroform, trizol, ethanol absolute, trypan blue, methanol, hexane, benzalkonium chloride, and pentanol are used and discarded according to the regulations and guidelines of NCHU’s Safety Center (full name: Center for Environmental Protection & Occupational Safety and Health). The experimental operations were all carried out in a fume hood, equipped with personal safety protection (see Protective equipment for details).

Protective Equipment

It is necessary to be equipped with qualified personal safety protection equipment (PPE) during the experiment and in qualified environments. We have gloves, goggles, and lab coats during the experiments.

Disposal

1. Biological waste is sterilized in an autoclave at 121°C for 20 minutes and discarded according to the rules of the NCHU Safety Center.
2. The disposal of chemicals is managed by university officials who transmits the waste to a licensed third party.

Safety Training

All the wet lab members of NCHU_Taichung would perform the experiments after taking proper laboratory training, including the use of a pipet, the safe way to execute microorganisms, the use of emergency instruments, and the right process while using high-speed centrifugation.

Bee Safety

When combining genetically modified organisms with insects such as bees, anti-escape systems need to be put in place. By increasing the number of obstacles (such as doors) or using bright light to lure them, the likelihood of a breakthrough will be greatly reduced. Also, we make sure there are no direct escape routes before starting work. However, reduced survival of individual bees will limit dispersal if bees accidentally escape. Nonetheless, as mentioned above, escapes of individual organisms should be reported to National Chung Hsing University.