Overview
Safety has been our team’s top priority throughout this project. As a high school team, we strictly followed iGEM’s safety policies during every stage of the project. We continuously reviewed, checked, and consulted on our safety practices as the project content and progress evolved, ensuring that all participating students, teachers, interviewees, and other stakeholders were engaged in a safe environment to the largest extent. We also hope to pass on our experience to support future iGEM teams in maintaining the high standards of safety.
Risk Identification
Biological Risks
We selected Chlamydomonas reinhardtii as our model organism. It is a BSL-1 organism with a well-established genetic background, no pathogenicity, and with very low biosafety risk.
The target gene used to synthesize our plasmid was the endogenous CPN60C gene from C. reinhardtii. This gene encodes a mitochondrial chaperone protein that assists in protein folding and maintaining cellular function, especially under pressure. It has no known toxic, allergenic, or pathogenic effects, and therefore poses very low biosafety risk.
For plasmid amplification, we used E. coli DH5-alpha, one of the most widely applied laboratory-engineered strains in molecular cloning. It is non-pathogenic, unable to survive or spread in natural environments, and is also classified as BSL-1, with very low biosafety risk.
Hygromycin and paromomycin were employed as antibiotics for selecting transformants. Both have long been used in transgenic research with C. reinhardtii and are supported by extensive experimental methods and data. Their use is limited strictly to laboratory screening. Since C. reinhardtii is non-pathogenic and does not spread resistance genes into the natural environment, the biosafety risk is very low and fully compliant with iGEM’s antibiotic resistance policy.
Chemical Risks
Our research involved the use of antibiotics, specifically paromomycin and hygromycin. Although applied at very low concentrations, the potential chemical risks associated with these substances were carefully considered.
PCR experiments required the use of dyes, which also pose certain chemical risks and were handled with caution.
Physical Risks
A laminar flow hood (clean bench) was used to maintain sterile conditions for some experimental steps. While iGEM discourages its use because it protects only the samples (not the operator or the environment), we employed it exclusively for non-pathogenic experiments with minimal risks. All such operations were performed under the direct supervision of laboratory instructors and consultants.
Security Risks
Our Human Practices activities included surveys and interviews with human participants. These were conducted in strict compliance with local regulations and iGEM policies, with particular attention to the involvement of minors and the protection of participant data privacy.
Our Strategies and Practices to Address Safety and Security
2.1 Safe Experimental Subjects, Materials, and Methods
As described above, our model organisms, target genes, and E. coli strains are all widely recognized as safe and extensively validated experimental materials. Theoretically, they pose no foreseeable risks to researchers, the public, or the laboratory environment.
We selected resistance screening over fluorescence screening, mainly because Chlamydomonas reinhardtii is photosynthetic and exhibits inherent fluorescence, which could interfere with the accuracy of fluorescence-based methods. Moreover, antibiotic screening is more economical, efficient, and widely adopted in related research. To ensure safety, we followed rigorous training protocols, enforced management systems, and implemented strict waste disposal procedures.
2.2 Comprehensive and Systematic Laboratory Safety Training
All team members involved in experimental work, including students, advisors and the instructor, were required to complete approximately six hours of comprehensive laboratory safety training. This consisted of two hours of online theoretical instruction and four hours of hands-on laboratory practice.
2.3 Strict Laboratory Safety Management System
- Only personnel who completed safety training and passed the assessment were permitted to enter the laboratory. Entry and exit were recorded with mandatory sign-in and sign-out.
- Personal protective equipment (PPE), including lab coats, gloves, and goggles, was required at all times.
- All experiments were conducted under the supervision and guidance of the laboratory instructor and/or the adult advisors.
- The purchase and storage of antibiotics and PCR dyes were strictly documented and maintained in a locked cabinet. The instructor’s authorization and instructions were required for any usage, and the minimum effective concentrations were strictly followed. PPE and N95 masks were mandatory during handling. Wastewater and media containing antibiotics were treated and disposed of separately as chemical waste.
- Culture media without antibiotics, even if posing no theoretical pathogenic risk, were thoroughly autoclaved prior to centralized collection and disposal.
2.4 Adequate and Effective Laboratory Safety Equipment
2.5 Strict Human Practices Regulations and Data Management
In conducting Human Practices, we fully complied with local laws and regulations as well as iGEM’s safety and ethics policies. Participation was entirely voluntary, with the option to withdraw at any time. For underage participants, guardian consent was required before involvement in surveys or interviews. All raw data were securely stored by a designated team member and accessible only to authorized team members and the P.I. No personal information collected was used for any commercial or non-commercial purpose outside the scope of the project.
Notes on Safety Considerations under Project Evolution
Our project underwent a Project Evolution (see Project Description for details), during which the target gene was changed from the endogenous Chlamydomonas reinhardtii gene LCIB to another endogenous gene, CPN60C. Like LCIB, CPN60C is non-pathogenic, non-toxic, and non-allergenic. Aside from their differences in sequence and biological function, both genes share the same research objectives, experimental methods, required laboratory conditions, and procedures. Therefore, this change had no impact on our safety measures.