Overview
Safety is the fundamental guarantee for the project of the Beijing Normal University 2025 iGEM BNU-China team. Based on the design and construction of the V-CHARGEs (VLP-Coupled High-efficiency ATP Re-Generating Enzyme system), we have always placed biosafety and standardized operations as our top priority, comprehensively preventing various risks to ensure personnel safety, protect the ecological environment, and secure the reliability and responsibility of future technological applications. Throughout the entire project cycle, we strictly adhered to iGEM safety guidelines, actively consulted biosafety experts, and integrated the concept of safety into every aspect of our work—including experimental design, material selection, operational procedures, and waste disposal—striving to achieve full control and zero hidden hazards during project advancement. This article systematically elaborates on the potential risks involved in this project and the comprehensive prevention and control measures that have been implemented.
Project Safety
Biosafety is the fundamental prerequisite for life science research, encompassing multiple dimensions such as standardized experimental procedures, biological material management, and environmental impact control. The BNU-China team, grounded in a systematic biosafety assessment, implements risk prevention and control strategies throughout the stages of project design, execution, and potential industrial translation, with the firm aim of eliminating risks of harm to laboratory personnel, environmental pollution, and threats to public health. Biosafety risks refer to the potential threats to human health, ecosystem balance, or agricultural production caused by biological factors, such as pathogens, pests, and invasive species. Therefore, it is necessary for us to identify and manage potential biosafety risks in our project to ensure the safety of experiment personnel, the laboratory environment, and public health, as well as to improve the safety and reliability of the experiments. This will lay a foundation for the future public and market adoption of our project results.
In the process of experiment design, through discussion, we have analyzed and identified potential biosafety risks in the following three aspects: our chassis organism, the experimental process, and the risks associated with the product reaching the public after future production.
The chassis organism used in this study is Escherichia coli BL21 (DE3), a routinely applied non-pathogenic laboratory engineering strain with no ability to spread in the environment. It meets Biosafety Level 1 (BSL-1) standards and poses no risk of biological contamination or ecological invasion. All expressed protein components—including polyphosphate kinase fused with the P22 bacteriophage Scaffold Protein (SlPPK-SP), P22 bacteriophage Coat Protein fused with SpyTag (CP-SpyTag), Uridine-Cytidine Kinase (UCK), γ-Glutamylcysteine Synthetase (γ-GCS), and Glutathione Synthetase (GS)—were evaluated by bioinformatics analysis and experimental validation, confirming that they are non-toxic, non-allergenic, and environmentally safe.
All chemicals involved in the experimental process (such as absolute ethanol, TEMED, and β-mercaptoethanol) are strictly managed in accordance with the Beijing Normal University (BNU) Regulations for the Management of Hazardous Chemicals in Laboratories. Hazardous chemicals are stored in classified categories under dual supervision; all reagent cabinets are locked and clearly labeled with storage conditions. Instrument operations such as centrifugation and pipetting follow standardized protocols, with equipment maintained and logged regularly, thereby minimizing risks of physical and chemical occupational hazards.
Looking toward future industrial applications, this project also demonstrates a solid foundation of safety: the final product is a polyphosphate kinase complex encapsulated within P22 Virus-Like Particles (P22-VLPs). These VLPs contain no nucleic acids, cannot self-replicate or infect, and are biodegradable under natural conditions, ensuring a high level of safety for both humans and the environment. The production strain, E. coli BL21 (DE3), is cultured entirely within a closed fermentation system and sterilized at high temperature upon completion of fermentation, eliminating the possibility of live-cell leakage or horizontal gene transfer. All raw and auxiliary materials are standard industrial-grade, and the purification process is mature. The overall workflow is green, low-carbon, and generates no hazardous waste, fully meeting the environmental protection and safety requirements of industrial-scale production.
Laboratory Safety
Beijing Normal University (BNU) has established a comprehensive and rigorous laboratory safety management system. All experiments in this project were conducted in laboratory environments meeting BSL-1 standards. All experimenters have completed university-level safety training and assessment, signed safety responsibility commitment forms, and received hands-on practical training. The laboratories are equipped with essential safety facilities, including biosafety cabinets, autoclaves, emergency showers, and eyewash stations. Daily management strictly complies with the Laboratory Safety Regulations for Higher Education Institutions and the Beijing Normal University (BNU) Laboratory Safety Management Measures.
Safety Regulations
Detailed Safety Management Regulations
1. Personnel Access and Training
(1) A laboratory safety access system is implemented: all personnel must pass examinations and sign commitment forms before entry;
(2) Regular safety training and practical re-training are conducted, with emphasis on high-risk operational skills.
2. Personal Protective Requirements
(1) Laboratory entry requires lab coats, closed footwear, masks, and gloves;
(2) When handling hazardous chemicals or biological samples, protective goggles or face shields must be additionally worn.
3. Standard Experimental Practices
(1) All biological experiments must be carried out inside a biosafety cabinet; open-bench operations are strictly prohibited;
(2) Pipetting must be performed with mechanical pipettors, and sharps must be disposed of immediately in designated containers.
4. Chemical and Biological Material Management
(1) Hazardous chemicals are managed under a “dual-person, dual-lock” system, with detailed logs for requisition and return;
(2) Biological samples must be recorded throughout storage, use, and disposal, ensuring full traceability.
5. Instrument and Equipment Use and Maintenance
(1) Large instruments require operation training and authorized access, with regular performance verification;
(2) Electrical devices must be used in designated locations away from water sources, and wiring is regularly inspected for hazards.
6. Waste Classification and Disposal
(1) Laboratory waste is strictly classified as biological, chemical, or physical, collected in separate, clearly labeled containers;
(2) All biological waste must be autoclaved prior to disposal.
7. Emergency Preparedness and Response
(1) Emergency contact numbers and response procedures are posted prominently in the laboratory;
(2) Regular emergency drills are organized for fire, chemical spills, and biological sample splashes.
8. Daily Management and Environmental Maintenance
(1) A rotating duty system is in place: before leaving, the last person must check the safety of water, electricity, gas, and windows/doors;
(2) Weekly safety inspection records are summarized, potential hazards rectified promptly, and documentation archived for reference.
Chemical Substances Safety
To ensure the safe management of chemicals in the laboratory, we have implemented a range of strict measures to safeguard team members and facilitate the smooth execution of experiments.
We strictly adhere to the laboratory safety management regulations of Beijing Normal University (BNU), ensuring that all experimental processes meet safety standards. All experiments involving potentially hazardous chemicals are conducted in dedicated chemical fume hoods to prevent the leakage of harmful gases and accidental exposure. Additionally, team members must wear appropriate Personal Protective Equipment (PPE) such as gloves, goggles, and lab coats when handling chemicals to minimize the risk of direct contact.
All chemical reagents used by Beijing Normal University (BNU) are purchased through legitimate channels. The quality of all chemical reagents is guaranteed.
Furthermore, we have conducted thorough inspections of the storage of all chemical reagents in the laboratory. All chemical reagents are categorized and stored according to their properties, with hazardous chemicals stored separately from other chemicals in designated, locked cabinets. We ensure that all chemical reagent containers are clearly labeled and accurately identified, and placed within the laboratory’s line of sight to prevent misuse or accidental exposure.
Moreover, we conduct regular safety inspections of the laboratory, including the storage conditions of chemical reagents, the operational status of chemical fume hoods, and the use of personal protective equipment.
Safety Training
Before the commencement of all experiments, the students responsible for experiments in our team have already undergone safety education on the use of equipment such as autoclaves, laminar flow hoods, biosafety cabinets, fume hoods, PCR machines, and centrifuges. They have also completed two semesters of basic chemistry lab courses. All students in our iGEM team have enhanced their awareness of lab safety and strictly adhere to the laboratory safety regulations during experiments.
With the help of our lab instructor, Dr. Hao Xiaoran, we conducted additional laboratory safety education jointly with the iGEM team from Beijing No.5 High School. The instructor taught the relevant regulations for biological and chemical operations. Experiment students are expected to conduct their experimental explorations according to these regulations, with the assistance and supervision of senior team members.
After completing the training for all members of the wet lab team, we passed the virtual simulation assessment for emergency response to safety incidents and signed and confirmed the lab safety agreement on-site.
Under this safety training, our experiments proceeded very smoothly. Over the course of several months of experimentation, no laboratory safety incidents occurred.
Safety in Human Practices
During the Human Practices activities, we fully guaranteed the personal and property safety of all the team members and participants. All members participating in the volunteer teaching activities have received systematic safety education. All students participating in the activity are fully guaranteed safety.
In the Human Practices, we ensured the personal privacy of every participant and their individual rights. Specifically, when collecting survey information, we committed to not using the gathered data for any other purposes. During interviews, if audio or video recording was required, we obtained prior consent from the interviewees. If we intended to publish the interview content online, we sought approval from the interviewees in advance for review. In essence, we made every effort to guarantee the personal privacy of our participants.