To comply with COVID-19 level 4 safety protocols (ban on gatherings of 5 or more in daytime and 3 or more after 6 PM) implemented by the Korean government, all experiments were completed before 6 PM in groups of at most 4 people. All wet-lab participants maintained a safety distance of at least 1 meter and were required to record their temperature (students with body temperature >37.5℃ or respiratory symptoms were advised to stay home) and complete an entry log (i.e. visiting time and date). Also, constant use of facial masks (N95 or higher), sanitization of participants and equipment, and ventilation were performed.
Our goal is to design a cancer-specific, cell-penetrating peptide for the efficient delivery of siRNA into cancer cells. The peptides will be synthesized by an automated peptide synthesizer. The effect of designed peptides will be analyzed in human lung cells and cancer cells. 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).
MRC5 (human lung cells) and A549 (human lung cancer 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.
Synthesized peptides will be tested on MRC5 and A549 for the analysis of penetration specificity, cytotoxicity, cargo transfection efficacy (intracellular distribution), and cell death. No risks could arise from these experiments.
If developed into a finished product, it will be used in the human body or the natural environment. Hence, this calls for a safety and security risk assessment that addresses the side effects of the synthesized peptides on human cells. No autonomous spread to the environment is expected.
The laboratory at the University of Suwon operates under level 1 (low risk) biosafety level. All team members received extensive safety and security training encompassing topics of lab access and rules, responsible individuals, differences between biosafety levels, biosafety equipment, disinfection and sterilization, emergency procedures, transport rules, physical biosecurity, personnel biosecurity, dual-use and experiments of concern, data biosecurity and cyber biosecurity, and chemicals, fire, and electrical safety. Open benches and biosafety cabinets along with personal protective equipment and waste management systems were used to handle biological materials.
- 1. Wear protective gear including latex gloves, masks, lab coats, closed-toe shoes, and long trousers to prevent chemical contamination on your body.
- 2. Never eat or drink in the laboratory.
- 3. Do not touch any lab equipment or chemicals until instructed to do so.
- 4. Tie back long hair.
- 5. Report any incidents to the instructor, no matter how minor.
- 6. Be fully aware of the building’s evacuation procedures.
- 7. Know the location of lab safety equipment, including fire extinguishers, safety showers, and first aid kits.
- 8. Be approved or trained by the supervisor of all equipment before first use.
- 9. Never leave an ongoing experiment unattended.
- 10. Never put unused chemicals back into the original container.
Risk Management Provisions
Tissue culture waste (culture medium) was inactivated for at least 2 hours in a solution of hypochlorite (10,000ppm) prior to disposal at the sink with an excess of water. Moreover, contaminated pipettes were placed in hypochlorite solution (2500ppm) overnight before disposal by incineration. Generally, the following guidance was used to address cell culture wastes.
- 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.
Compliance to iGEM Safety/Security Policies
- 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 of 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.