Safety is one of the core priorities of our project. All experiments were conducted following biosafety regulations and under the supervision of qualified instructors at National Chung Hsing University (NCHU). Our goal is to ensure that every stage of our work—from design to experiment—is performed responsibly, with no risk to people, other organisms, or the environment.
All wet-lab members completed:
This training strengthened our team's understanding of risk assessment and laboratory containment principles, ensuring all work met the highest safety standards.
All laboratory work was performed in a BSL-1 (Biosafety Level 1) facility certified by NCHU, which is suitable for handling non-pathogenic microorganisms.
Facility Equipment:
We carefully selected organisms and materials with established safety profiles to minimize any potential risks throughout our research.
| Organism | Classification | Description & Safety Information |
|---|---|---|
| Kluyveromyces marxianus | BSL-1, GRAS | The primary host organism used in our project. Recognized by the U.S. FDA as Generally Regarded as Safe (GRAS) and widely used in the food and fermentation industries. |
| Escherichia coli DH5α | BSL-1 | Used for plasmid amplification and cloning. This non-pathogenic strain is safe for standard molecular biology use. |
| Gene Sources | DNA sequences only | The A2 β-casein gene (CSN2) from Bos taurus / Bos grunniens, and carbon fixation genes (RuBisCO, PRK) were synthesized in vitro and do not involve live animal or plant materials. |
Important: No pathogenic microorganisms, toxins, or antibiotic-resistance genes were used in any part of the project.
To ensure biosafety and environmental containment, we implemented multiple strategic approaches:
Engineered Kill Switch
Our engineered yeast vector includes a genetic kill switch that activates autophagy and cell death under specific conditions, preventing uncontrolled survival outside the laboratory.
Targeted Genomic Integration
All foreign genes were integrated directly into the yeast genome rather than maintained on plasmids, enhancing genetic stability and minimizing horizontal gene transfer risk.
Physical Containment
All experiments were performed in sealed culture systems with biological waste sterilized via autoclaving before disposal.
Regulatory Compliance
Strict adherence to Taiwan CDC's Laboratory Biosafety Management Guidelines, ensuring all procedures met national standards for microbial research.
Our comprehensive safety protocols included:
Our project does not use live animals or higher organisms. Although the β-casein gene originates from cattle or yak, all work uses synthetic DNA sequences obtained from databases such as NCBI and synthesized commercially.
No animal experimentation or tissue collection is involved. The project fully complies with iGEM's animal use policy and NCHU's institutional biosafety guidelines.
Safety is not just a requirement—it's our fundamental commitment to responsible scientific innovation.