Ethic and Safety

The experiments were carried out in the laboratory (6th floor) of the Medical Genetics Research Center of the School of Life Sciences, Central South University. Our laboratory is classified as BSL - II, complying with the "General Requirements for Laboratory Biosafety" [1] and the "Regulations on the Safety Management of Pathogenic Microorganism Laboratories" [2]. The laboratory layout is reasonable and well - organized.

Experimental environment and storage environment

1. Laboratory Environment

Figure: Laboratory Environment Overview

2. Cell Room—Biological Safety Cabinets, Incubators, Gas Cylinders

Figure: Biological Safety Cabinets in Cell Culture Room

Figure: Cell Culture Incubators

Figure: Gas Cylinders with Safety Measures

3. Fire Extinguisher

Figure: Fire Extinguisher and Emergency Safety Equipment

Before entering the laboratory, all members must undergo laboratory safety training and laboratory skills training. Safety training includes health and safety protection, instrument use safety, chemical use safety, fire safety, etc. Skills training includes cell culture and passage, plasmid expansion and transfection, dual - luciferase reporter gene detection, and WB. Members who have not undergone safety training and skills training are not allowed to enter the experiment. This is not only to ensure the validity of experimental data but also to ensure the safety of members and the laboratory.

Figure：The above picture shows the online training website: https://labexam.xauat.edu.cn/redir.php?catalog_id = 123&page = 1

We strictly abide by all specific guidelines regarding biosafety when using the laboratory. The following are some of the laboratory guidelines:

Equipment	Hazards to avoid	Safety features
Lab coat, isolation gown, jumpsuit	Contaminating clothes	Open at the back and worn over daily clothing.
Plastic apron	Contaminating clothes	Waterproof.
Shoes and socks	Collision and splashing	Toes are covered.
Goggles	Collision and splashing	Anti - collision lenses (must have vision correction or wear vision correction glasses externally), with side shields.
Safety glasses	Collision	Anti - collision lenses (must have vision correction), with side shields.
Face shield	Collision and splashing	Covers the entire face and is easy to remove in case of an accident.
Gas mask	Inhaling aerogels	Designed to include disposable, full-face or half-face air-purifying, full-face or powered air-purifying respirators with a hood, and supplied-air gas masks.
Gloves	Direct contact with microorganisms	Disposable latex, vinyl resin, or nitrile-based protective gloves approved by microbiology.

The waste in the biological laboratory mainly includes infectious waste, pathological waste, and sharp waste.

· Infectious waste: Medical waste that carries pathogenic microorganisms and has the risk of spreading infectious diseases.

· Pathological waste: Human waste generated during diagnosis and treatment and the corpses of medical experimental animals, etc.

· Sharp waste: Discarded medical sharps that can stab or cut the human body. Safety disposal precautions:

1. Collect and transfer biological waste in a timely manner.

2. Store biological waste separately from chemical waste and household waste.

3. Equip the laboratory with biological waste trash cans (with special plastic bags for biological waste inside) and paste special labels.

4. Sharps such as blades and pipette tips should be placed in sharps containers or puncture - resistant cardboard boxes and then put into special plastic bags for biological waste and labeled when being stored.

5. After the animal experiment, the animal corpses and tissues should be harmlessly treated, and the waste should be thoroughly sterilized before disposal.

6. Biological waste involving pathogenic microorganisms or other bacteria must be treated by high - temperature and high - pressure sterilization or chemical soaking, and then finally disposed of by a qualified company.

7. Implement traceability tracking for the disposal of highly pathogenic biological material waste.

Figure：Household trash can and contaminated trash can

Self-checked entry	Answers
1. Did the team make a contribution to biosafety and/or biosecurity?	Yes, we provided a reference for subsequent experimental steams about determining optimal transfer plasmid combination of spilt lactic acid sensor and inactivated TEV protease system in the circumstance of applying human embryonic kidney 293T cells as target host cells.
2. Is their contribution well - characterized and/or well - validated?	Yes, we modeled and predicted the dynamic responses of 8 fusion protein configurations through testing the sensitivity, sLOx production, leakage, and response speed. We screened out the best transfer plasmid through experiments and verified the function of the system in degrading extracellular lactic acid through cell experiments.
3. Did the team build upon existing knowledge, understanding, tools or approaches?	The project was based on discussions with our teachers and researchers who have been working in the field of immunotherapy. The research originated from the challenges they encountered during their work. The synthetic biology techniques are derived from previous literature, iGEM teams, and our lab's work over the past several years.
4. In addition to applied safety work, has the team managed any risks from their project appropriately?	Yes, we considered the possible risks in the project, verified and avoided them through experiments, and discussed them with experts and peers.
5. Has the team addressed the use of synthetic biology beyond the iGEM competition?	Yes, our project provides reference data on the therapeutic effect of cell therapy on solid tumors. Meanwhile, it's a breakthrough in publicity regarding the public perception barrier of cell therapy, which has not been put into application. The popular science of modified cells and protein engineering in the project is a good way for beginners in synthetic biology to get started.

At the CCiC conference, we attended a lecture on biosafety and ethics, where experts emphasized their importance in biological experiments.

One key insight was, “When you're unsure if you've made a mistake, discuss it with your peers.” Inspired by this, we organized a seminar on biosafety and ethics with other IGEMers from China. Participants shared their projects, followed by suggestions for improving biosafety safeguards.