With the advancement of synthetic biology, safety has become a core concern and a vital concept in iGEM. LINKS-China is dedicated to integrating safety into every aspect of our project, ensuring that each step aligns with responsible research practices.

To maintain high safety standards, we have implemented multiple safety measures throughout our project. In our design, careful consideration is given to ensure biosafety, and our experiments are conducted under strict laboratory safety protocols. All team members are thoroughly trained and adhere to lab safety guidelines. In our Human Practices, we prioritize confidentiality by establishing a privacy policy for any collected personal information. Furthermore, we ensure that our educational activities comply with established safety regulations, emphasizing the importance of safety in synthetic biology research.

We first considered the potential risks during the brainstorming and experimental design phases, identifying possible hazards before proceeding. This allowed us to proactively address and mitigate those risks, ensuring compliance with safety protocols throughout the project.

As outlined in our project design, we prioritize the safety and well-being of all participants and the environment throughout the research process. Our laboratory operates under stringent biosafety protocols and is classified as a Level 2 containment facility, ensuring that all biological materials are handled with care. We utilize biosafety cabinets for handling our engineered organisms, distinguishing them from laminar flow hoods to maintain a sterile environment.

To mitigate potential risks associated with our project, we have implemented comprehensive safety measures, including:

a. Training and Education: All team members undergo rigorous training in laboratory safety, with a focus on proper handling techniques for biological materials and emergency response protocols.

b. Incident Reporting: We have an incident reporting system in place to record and address any accidents, fostering a culture of transparency and accountability within our team.

Our Goal:

Our project involves engineering Escherichia coli (DH5-alpha, BL21, SHuffle) and Pichia pastoris yeast to produce fusion proteins. These fusion proteins will consist of binding domains for common materials in first-aid kits, proteolytic cleavage tags, and human antimicrobial peptides (AMPs). The peptides will be extracted and purified from the engineered bacteria/yeast. The genes encoding the AMPs and cleavage tags are derived from the human genome.

By integrating strict safety practices into our research design, our goal is to ensure that our innovative solution not only addresses critical medical challenges but is also conducted responsibly and ethically.

Risks:

- Handling Staphylococcus aureus: Opening culture plates containing Staphylococcus aureus outside the biosafety cabinet or spilling it onto the biosafety cabinet surface.
- Bacterial Use: While we frequently use Escherichia coli strains (DH5-alpha, BL21, SHuffle) and Pichia pastoris (both listed as safe organisms), Staphylococcus aureus is a BSL-2 pathogen.
- Improper Handling of Staphylococcus aureus: Can lead to food poisoning or skin infections if mishandled.
- Fermentation Outside Biosafety Cabinets: Large-scale fermentation and lysis of bacteria, particularly when handling potentially hazardous strains.
- Spillage of Fermentation Broth: Spillage of bacterial fermentation broth can lead to leaks and contamination.

Safety Measures:

- Biosafety Level 2 Protocols: Work involving Staphylococcus aureus must be conducted inside a biosafety cabinet.
- Proper Use of Biosafety Cabinets: Ensure the correct use of biosafety cabinets for all handling of hazardous materials.
- Fermentation of Non-Hazardous Strains: Large-scale fermentation, lysis, and handling of bacteria outside the laminar flow hood will only involve the safe-listed strains, E. coli and P. pastoris, and not Staphylococcus aureus.
- Personal Protective Equipment (PPE): Minimize direct contact with the bacteria by wearing gloves and masks during inoculation and handling. Use alcohol for flame sterilization before handling cultures.

- Safe Handling of Bacterial Samples: Securely cap centrifuge tubes to prevent spillage of fermentation broth containing bacteria. After centrifugation, supernatants will be disposed of in a sterile environment, disinfected with bleach, and safely discarded.
- Sterilization: Clean the needle of the homogenizer thoroughly with 100% alcohol after use to prevent contamination.
By adhering to these measures, we ensure safe handling of all biological materials, minimize risks, and maintain a controlled research environment.

Our workspace operates under a Level 2 containment system to ensure a safe environment for handling biological materials. The laboratory is classified as Biosafety Level 1 (BSL-1), which indicates that all materials pose minimal to no risk to personal health. The lab is divided into three distinct areas: work area, rest area, and laboratory area. The laboratory area is further separated into contaminated and non-contaminated zones.

We use biosafety cabinets for all tasks involving biological materials, as they provide the necessary protection. Unlike laminar flow hoods or clean benches, which are not recommended for handling potentially hazardous materials, biosafety cabinets are designed to contain dangerous agents.

Risks

- Chemical Hazards: The laboratory frequently uses chemicals such as disinfectants, fixatives, or reagents, which may be toxic, corrosive, or carcinogenic.
- Sharp Objects: Needles, scalpels, and broken glass can cause cuts or punctures, potentially leading to infections or exposure to harmful substances.
- Biohazardous Waste: Improper disposal of biohazardous materials, such as contaminated culture plates, can lead to the spread of infectious agents.
- Fire and Explosion Hazards: Flammable chemicals, open flames from Bunsen burners, or faulty equipment may cause fires or explosions.

Safety Measures:

- Centrifuge Use: Large instruments such as floor-standing centrifuges are balanced with precision, with a tolerance of less than 0.1 grams. Users are instructed not to leave the centrifuge until it has reached maximum speed and is operating safely.
- Lab Regulations: Strict adherence to lab protocols is enforced, including securing hair and keeping personal items out of the work area.
- Agarose Gel Electrophoresis: This procedure is confined to the contamination zone, using designated pipettes and conical flasks. Solutions containing microorganisms are collected in specific containers and treated with bleach (sodium hypochlorite) before safe disposal. Petri dishes and other contaminated materials are sterilized using an autoclave to ensure no viable microorganisms are released from the lab.
- Biohazard Disposal: Appropriate biohazard protocols are followed, including sterilizing waste in autoclaves and disposing of it in biohazard bags.
- Storage and Fire Safety: Flammable chemicals are stored appropriately, and flame-resistant lab coats are worn. Fire extinguishers and emergency exits are kept easily accessible.
- Equipment Safety Training: Proper training in the correct and safe use of laboratory equipment is mandatory. Passing a safety exam is required before participating in experiments.
- Prohibited Activities: Strict guidelines prohibit eating, drinking, using phones, and removing laboratory waste or materials without following proper disposal procedures.
- Protein Electrophoresis: Special care is taken to avoid contact with SDS during protein electrophoresis to prevent skin exposure.

- Gel Contamination Area: Masks and gloves are worn in the gel contamination area to avoid exposure to toxic substances and prevent skin contact with nucleic acid stains.

- E. coli Handling: When handling E. coli outside the laminar flow hood, strict protocols are followed, including wearing masks, using alcohol, and open flames, to avoid human contact with E. coli.

By adhering to these protocols, we ensure a minimal risk to personal health, while the laboratory remains organized into separate work, rest, and experimental zones, with designated contamination and non-contamination areas.

Our project aims to address a critical healthcare challenge: the risk of wound infections in areas with limited medical resources. In high-stress environments—such as battlefields, remote communities, impoverished regions, and outdoor settings—untreated wounds can quickly become life-threatening due to inadequate care and the growing threat of antibiotic resistance.

To tackle this issue, we are developing a first-aid kit that incorporates antimicrobial peptides, providing an immediate and powerful solution that surpasses traditional treatments. This innovative approach not only reduces the risk of severe infections but also alleviates the burden on healthcare systems by decreasing reliance on antibiotics, making a significant contribution to the global fight against antibiotic resistance. Our solution empowers individuals and communities to manage medical emergencies effectively, improving health outcomes, reducing healthcare costs, and enhancing societal resilience during medical crises.

Risks:

- Legal Compliance and Informed Consent: Ensuring compliance with legal regulations and obtaining informed consent from participants.
- Privacy Infringement: Safeguarding participants' privacy during interviews and outreach activities.
- Incident Reporting and Follow-up: Inadequate systems to track and follow up on incidents during the project.
- Lack of Professionalism: Providing inaccurate or misleading information to those receiving education or participating in outreach.

Safety Measures:

- Legal Compliance and Consent: Strictly adhere to legal regulations, ensuring participants' right to informed consent. Participants will be fully briefed on the interview's purpose and will have the right to decline answering any questions. A detailed activity plan will be drafted and provided to participants in advance for review and approval before proceeding with interviews.
- Privacy Protection: During outreach and educational activities, informed consent will be obtained from all participants, with full respect for their privacy and rights, ensuring they feel valued and respected.
- Incident Reporting: A robust reporting system will be established to document and address any incidents promptly, fostering a culture of safety and accountability within the team.
- Professionalism: Ensure that all team members receive proper training and have a strong sense of responsibility before engaging in interviews or educational activities to avoid the dissemination of incorrect information.

Through these comprehensive measures, we uphold the values of responsibility, transparency, and ethical engagement as outlined by iGEM. Our commitment to safety extends beyond laboratory practices, encompassing all our interactions with the community to ensure that our project remains innovative and aligned with the highest standards of human practice.