Policy compliance
Throughout the implementation of our project, we have always prioritized policy compliance and safety as the core principle. We strictly adhere to all policy requirements of iGEM as well as national policy regulations to ensure the safety and compliance of the project.
Safety in Our Strains
The Escherichia coli strains TOP10, DH5α, BL21(DE3) and Nissle 1917(EcN) selected as chassis for our project are all included in the iGEM White List . These E. coli strains have been subject to comprehensive, rigorous safety assessments, confirming they exhibit no pathogenicity, produce no toxic metabolites, and pose no adverse effects on human health nor risk of disrupting the natural microbial ecosystem. Throughout the experimental process, we strictly use only strains listed in the iGEM White List, prohibit using any unapproved strains or those with potential safety risks, and thereby ensure compliance with iGEM's policies regarding strain usage.
Furthermore, the toxin/antitoxin genes ccdB and ccdA used in our project were obtained via PCR from a plasmid of Salmonella (obtained from Professor Baojun Wang's lab). Salmonella is an intestinal pathogenic bacterium commonly found in nature, capable of causing various diseases such as typhoid fever, paratyphoid fever, and food poisoning. Prior to use, we submitted a check-in form and obtained permission from the Safety Committee of iGEM. During the experimental process, we strictly followed standard laboratory operating protocols (SOPs), adhered to appropriate personal protective measures (PPMs), and strictly prohibited the unauthorized removal of bacterial strains from the laboratory. Additionally, after use, Salmonella cultures and PCR waste liquids were sterilized with alkaline solutions, then properly disposed of.
Figure 1. Alkaline solutions for sterilization
Specifications for Experimental Operations
General Safety Guidelines
All team members should complete systematic laboratory safety training before conducting experiments. The training covers key knowledge and skills including chemical usage, waste disposal, experimental ethics, experimental record, item labeling, operation of safety equipment and emergency response. This ensures all laboratory members understand and subsequently comply with the "Laboratory Code of Conduct", upholding experimental safety at its source.
Unauthorized acquisition or removal of laboratory items, bacterial strains, and other materials is strictly prohibited to maintain the laboratory's safety order.
All personnel involved in experiments must gain a comprehensive and in-depth understanding of the experimental procedures, potential risks and precautions before experiments, so as to ensure the accuracy and safety of experimental operations.
Figure 2. Laboratory safety training of HiZJU-China
During experiments, all laboratory personnel should wear standardized rubber gloves throughout the process to avoid direct skin contact with experimental materials, thereby minimizing the risks of infection and cross-contamination.
All operations that involve viable microorganisms should be performed inside a super clean bench. The protective mechanism of the clean bench effectively prevents microorganisms from leaking into the laboratory environment. Besides, after using the biosafety cabinet, the UV lamp should be promptly turned on for sterilization for at least 30 minutes.
Figure 3. UV mode for sterilizing the clean bench after use
When using electrical equipment such as microwaves, personnel must be on-site for supervision throughout the operation to prevent safety accidents like fires caused by unattended equipment.
All liquid and solid wastes may containing living organisms should undergo strict sterilization treatment before subsequent disposal, so as to prevent the spread of harmful microorganisms.
After the completion of daily experiments, the last person to leave the laboratory must carefully check and turn off heating equipment including water baths, autoclaves and microwaves, and lock doors to eliminate potential safety hazards.
Safety Guidelines for Specific Strain Manipulation
For operations involving various E.coli, we have implemented additional measures:
1. Using dedicated experimental instruments and consumables to avoid cross-contamination between different strains;
2. Thoroughly disinfecting workbenches with 75% ethanol before and after experiments to create a safe experimental environment;
3. Maintaining detailed records of relevant experimental data, including strain usage, experimental conditions and results, to facilitate subsequent traceability and analysis.
For operations involving Salmonella, we strictly implement enhanced biosafety measures in line with biosafety level 2 (BSL-2) requirements. Salmonella is used solely for PCR, with no other experimental applications. After successfully obtaining the antitoxin gene ccdA and toxin gene ccdB, Salmonella cultures and PCR waste were sterilized using alkaline solutions and properly disposed of, thereby fundamentally preventing Salmonella from escaping into the environment and causing harm.
Management of Project Outcomes
All outcomes generated by our project, including experimental data, modified bacterial strains, and related products, are strictly confined within the laboratory. At all times, the release or use of project outputs, especially genetically modified organisms (GMOs) and their products, outside the laboratory is strictly prohibited, including during the competition period and at official events such as the iGEM Jamboree. In case of special circumstances, we will submit an application to the iGEM Safety and Security Committee and other relevant safety organizations in advance, providing detailed information on the nature, purpose, and safety assurance measures of the outcomes. Only after obtaining explicit approval will we proceed in accordance with the specified procedures, ensuring full compliance with iGEM's "No Release" policy.
Furthermore, we strictly respect others' intellectual property rights (IPR). All parts, research outcomes, and academic works used in experiments are clearly cited and attributed, fundamentally avoiding potential IPR disputes. Additionally, we will not disclose any other unpublished research outcomes or data from our team without authorization. By upholding data security and cybersecurity, we contribute to fostering a sound environment for knowledge protection.
Compliance with National Policies
Throughout our project implementation, we have closely monitored compliance with relevant national policies and regulations. In terms of pathogenic microorganism management, the E. coli strains used are non-pathogenic, fully complying with the Regulation on the Bio-safety Management of Pathogenic Microbe Labs. Should our project involve medical-related biological research in subsequent stages, we will adhere strictly to the Measures for the Ethical Review of Biomedical Research Involving Humans to fully protect research participants' rights and interests and ensure compliance with ethical standards. In the event of drug development or similar relevant efforts, we will also act strictly in accordance with policies and regulations such as the Measures for the Administration of Drug Registration issued by the National Medical Products Administration (NMPA). These measures ensure the legality and standardization of all project phases, and reflect our proactive fulfillment of obligations to comply with national policies.
Figure 4. Relevant national laws and regulations for project compliance (Left: Regulations on the Biosafety Management of Pathogenic Microorganism Laboratories; Right: Drug Administration Law of the People's Republic of China)
References
[1] The State Council of the People's Republic of China. (2004). Regulations on the Biosafety Administration of Pathogenic Microorganism Laboratories (Order No. 424 of the State Council. Revised by the Decision of the State Council on Amending Some Administrative Regulations on February 6, 2016, March 19, 2018, and December 6, 2024).
[2] National Health and Family Planning Commission of the People's Republic of China. (2016). Measures for the ethical review of biomedical research involving humans (Order No. 11). http://www.nhc.gov.cn/qjjys/s3581/201610/84b33b81d8e747eaaf048f68b174f6c2.shtml
[3] National Medical Products Administration. (2020). Measures for the administration of drug registration (Order No. 27). http://www.gov.cn/zhengce/zhengceku/2020-03/31/content_5497316.htm
[4] National People's Congress of the People's Republic of China. (2019). Drug Administration Law of the People's Republic of China. http://www.npc.gov.cn/npc/c30834/201908/6d97cf5596be4c6d93d5a369dcf905a4.shtml