Regarding COVID-19 Virus

With the ban of restrictions implemented in private gatherings on April 18th, we initiated our lab meeting in groups of 8 people beginning on July 3rd. However, as the guideline maintained the mask mandate policy, all wet-lab participants wore facial masks throughout the experiment. We have also maintained at least 1 meter social distancing, following the Ministry of Health and Welfare (MOHW) recommendation.

Lab Safety Rules

We have observed and followed all the general/basic laboratory safety rules under the guidance of Prof. Dr. Lee, as enumerated below:

1. Took note of the location of fire extinguishers, safety showers, and emergency exits.
2. Food and beverage consumption was strictly prohibited in the laboratory.
3. Everyone wore protective gloves, close-toed shoes, and lab coats in order to prevent skin & eye contact from chemical contamination.
4. Students tied their hair up in order to minimise chemical exposure.
5. Students all acted in a mature, responsible manner without horseplay.
6. We followed disclosure guidelines provided in "Standards for Disposal of Household Waste and Designated Waste."
7. We only left the laboratory when experiments were completed and no dangerous procedure was ongoing.
8. We never reused chemicals or containers.
9. We were informed to report minor accidents and mistakes.
10. We were guided thoroughly by the instructor before equipment use.

Project Safety

Our goal is to design a TFAM-DNA complex to ensure the protection of data in DNA. Please refer to the provisions below to review our team’s compliance with protocols issued by iGEM’s Safety and Security Committee and the South Korean government (Laboratory Animal Act).

• A172 (human brain cells), MCF7 (human breast cells), MKN45 (human stomach cells), and A549 (human lung cells) were purchased, posing no risk to the community or the environment if escaped from the lab. A few chemicals, including ethanol, cell culture media, and dimethyl sulfoxide (DMSO), may pose a hazard if improperly treated.
• A low concentration of nickel was used to isolate the TFAM protein from the solution, posing no risk to the environment when disposed of from the lab.
• Imidazole was used to separate the protein from ni, which does not pose serious harm to humans when exposed and to the environment when disposed of.

• For A172 (human brain cells), MCF7 (human breast cells), MKN45 (human stomach cells), and A549 (human lung cells), agarose gel was done for the selection of human cDNA to be used for PCR and to be put into a vector. No risks have arisen from this activity.
• The selected human cDNA was ligated to collect the plasmid inserted E. coli. Heat shock was provided to the e-coli containing tube to ligate the DNA. No risks have arisen from this activity.
• UV light and H2O2 were provided to the pSmile-TFAM complex to test whether adding TFAM enhances the stability of the data (pSmile). However, through providing regulated UV light and H2O2, the potential danger posed to humans and the environment was negotiated.

• If developed into a finished product, it will be used on a large scale. Hence, this calls for a safety and security risk assessment that addresses the risk when used in a grander scale. No autonomous spread to the environment is expected.

Risk Management Provisions

Tissue culture waste (culture medium) was deactivated for at least two hours in a solution of hypochlorite (10,000 ppm) prior to disposal at the sink with an excess of water. Moreover, contaminated pipettes were placed in hypochlorite solution (2500 ppm) overnight before disposal by incineration. Generally, the following guidance was used to address cell culture wastes, as indicated below:

1. Autoclave or disinfect the contaminated culture and associated media and dispose of it.
2. Thoroughly clean and disinfect all incubators, centrifuges, refrigerators, microscope stages, and any other equipment, including pipettors, that may have been in contact with the cell culture wastes.
3. All media, media components, and other reagents used for the contaminated cell lines must be discarded, and all other cell lines in use within the laboratory should be quarantined and tested for mycoplasma to detect any spread of contamination.
4. The experiment took place at the University of Suwon, a BSL1 laboratory. In order to ensure a safe experiment, our team implemented several safety measures, such as utilizing biosafety equipment, receiving project-specific safety and security training, and creating a thorough communication plan.
5. Personal protective equipment such as lab coats or gloves was provided to all members and the instructor when entering the lab.
6. The participating members were educated on the subjects such as emergency procedures, lab access, and disinfection prior to experiments to ensure the safety of each other and the supervisor.

Compliance to iGEM Safety/Security Procedures

1. “Do Not Release” Policy: Our team’s wet lab projects (i.e., source and/or product of genetically engineered organisms) were not deployed outside the lab and were limited to the experimental phase.
2. “No Human Experimentation” Policy: Our team’s project does not involve human testing or direct contact with humans with no requirements for human samples (swabs, blood, etc.).
3. “Whitelist” Policy: Our team’s project does not involve any animals, parts, or activities outside the breadth of the Whitelist or (higher-order) vertebrates -- with all parts and organisms obtained from trusted commercial and institutional suppliers.
4. “Human Subjects Research” Policy: All social science research (surveys and interviews as public engagement) was completed under stringent oversight to guarantee ethical and responsible research issued by the Institutional Review Board on informed consent, privacy and data protection, etc.