Safety

Safety is the foundation of our project and is integrated into every stage—from AI-based design to wet-lab experimentation and waste management. All work was conducted in Biosafety Level 1 (BSL-1) laboratories at National Tsing Hua University (NTHU) under the supervision of Assistant Professor Tsun-Hsien Hsiao (Department of Post-Baccalaureate Medicine).

We strictly follow the biosafety regulations of the NTHU Environmental Safety Center and the iGEM Safety and Security Policies. All members completed official Biosafety Training for Biological Laboratories, which covered laboratory conduct, waste handling, PPE use, and emergency procedures.

By combining responsible supervision, safe-by-design principles, and thorough training, our team ensures that our work remains safe for researchers, the public, and the environment.

Our project combines AI-assisted sequence design and directed evolution to develop non-hazardous and optimized genetic constructs. All sequences generated in Dry Lab were screened to eliminate any similarity to pathogenic or toxin-related genes.

Experimental validation was conducted in non-pathogenic E. coli DH10B, a cloning-optimized strain suitable for BSL-1 conditions. The project does not involve human or animal cell lines, pathogenic organisms, or environmental release.

By integrating computational design, rational screening, and laboratory containment, we ensure a safe, traceable, and biosafety-compliant synthetic biology workflow.

Plasmid delivery was performed by electroporation into E. coli DH10B using standard molecular cloning techniques. The plasmids, designed by our team and synthesized by IDT, contain a GFP reporter and were used solely for in-lab testing. Antibiotic selection was applied only for screening.

All procedures followed NTHU biosafety and waste disposal protocols, including autoclaving of all biological materials. No genetic material was transferred to other species or eukaryotic systems, ensuring complete laboratory containment.

All experiments were performed in BSL-1 facilities equipped with laminar flow hoods, centrifuges with safety locks, and sterilization systems. Members wore PPE at all times, and food or phone use was prohibited. Each sample and plasmid was properly labeled for traceability.

New members received hands-on training under the supervision of Assistant Professor Tsun-Hsien Hsiao and senior teammates before independent operation. At least two people were present during experimental work, and regular safety inspections were carried out.

Emergency tools such as eyewash stations, spill kits, and fire extinguishers were available, and all members were familiar with emergency response procedures.

Our team operates under NTHU biosafety regulations and international guidelines from the World Health Organization (WHO).

All members completed the NTHU Environmental Safety Center's Biosafety Training, which included waste disposal, PPE, chemical handling, and emergency response. Only members who passed both theoretical and practical training were authorized to perform experiments independently.

By adhering to institutional standards and ensuring thorough education, we maintain a culture of responsibility and biosafety awareness.

All biological and chemical wastes were treated according to NTHU's biosafety protocols.

Biological waste (cultures, tips, media) was autoclaved at 121 °C for 30 min, sealed, and collected by the Environmental Safety Center.

Chemical waste (solvents, antibiotics, dyes) was stored in labeled containers and removed by certified contractors.

Workspaces were disinfected with 70% ethanol or sodium hypochlorite, and sterilization logs were maintained. Regular inspection of autoclaves and safety devices ensured proper function and compliance.

All instruments were used only by trained members and supervised by Assistant Professor Tsun-Hsien Hsiao. Regular inspection, calibration, and maintenance logs were maintained.

Key equipment and safety practices include:

• Centrifuges: Samples balanced; lids locked during operation.
• Electrophoresis: Low voltage, safety covers; blue-light imaging replaced UV.
• Incubators: Temperature-calibrated and cleaned regularly; spills disinfected.
• Laminar Flow Hoods: Surfaces disinfected before/after use; UV lights off during operation.
• Autoclaves: Operated only by trained users.
• Micropipettes: Sterile tips; routine calibration.

The team also adopted safer reagents such as SYBR Safe instead of ethidium bromide to further reduce risks.

Although our experiments primarily involved low-risk reagents, we followed NTHU chemical safety regulations and maintained a detailed chemical inventory.

All reagents were clearly labeled and stored according to hazard classification. Safety Data Sheets (SDS) were available digitally and in print, and members reviewed them before handling chemicals. Acids, bases, and flammable materials were stored separately, while antibiotics and dyes were sealed in labeled containers.

Hazardous reagents (e.g., β-mercaptoethanol, EtBr) were collected in dedicated containers for removal by the NTHU Environmental Safety Center. The team also prioritized safer alternatives and eco-friendly reagents whenever possible.

From computational design to wet-lab experiments, every step of our project followed the principles of biosafety, containment, and responsible research. By adhering to institutional guidelines, maintaining continuous supervision, and fostering a strong safety culture, our team ensures that our synthetic biology work remains safe, ethical, and sustainable.