Parts Safety

None of the parts we used were hazardous. We used DH5-alpha competent cells, BL21DE3 competent cells, and BY4741 yeast strain for our experiments this year. Still, to prevent bacteria from entering the environment, we thoroughly autoclaved or bleached E.coli plates, yeast plates, tips, and tubes before disposal.

Our plasmids do not present any risk of entering other organisms. Specifically, the cloned pGal1,10-IsPETase plasmids can only be transformed into yeast and bacteria since the plasmids only have 2μ ORI for yeast DNA replication and ORI for bacterial DNA replication. Similarly, the cloned T7-IsPETase plasmids can only be transformed into bacteria since the plasmids only have ORI for bacterial DNA replication. Furthermore, bacteria and yeast will not pick up on either plasmids without specific selection on the plates.

The parts our team created will not present any risks if they somehow entered other organisms. This is because of our promoters, pGal1,10-IsPETaseS121E /D186H/R280A plasmids and T7-IsPETaseS121E/D186H/R280A, only being inducible in the presence of galactose and IPTG(Isopropyl β-D-1-thiogalactopyranoside) respectively. Thus, they will not be able to express in other organisms without these inducible compounds.

Lab & Project Safety

We did not do any activity prohibited in the iGEM competition, and all our work is covered by the White list. In our experiment, we only used permitted yeast and bacteria, with no other organisms or hazardous chemicals being used in the project. We also did not use artificial intelligence in the project.

In Kang Chiao International School Xiugang Campus, we used a level 1 standard microbiological lab consisting of an open bench, waste containers, and a chemical fume hood. There was also a chemical waste container that separated organic and inorganic waste. Moreover, there were separate biohazard waste containers, one for bacteria and yeast plates, and another for tips and Eppendorf tubes, to avoid risks of bio-waste contamination. In addition, the Esco Airstream Laminar hood in the laboratory helped us avoid contamination during bacteria and yeast inoculation.

Safety Training

All of our team members received safety training, including familiarization with lab access procedures, microbial techniques, personal biosecurity, biosafety equipment usage, and emergency protocols. We ensured that all team members undertook experiments safely and responsibly. We also implemented biosafety and biosecurity measures, including medical surveillance, data access controls, and the use of Personal Protective Equipment (PPE) to mitigate risks. The team was also supervised by Ms. Kuo, an experienced researcher and teacher in the field of microbiology, when handling organisms and other chemicals. In addition, iGEM ambassador Sabrina Yao and three university labs have checked that the KCIS laboratory meets all the safety requirements.

Our team referred to guidelines presented by the Taiwan Centers for Disease Control (CDC) and the Taiwanese Ministry of Education to ensure our safety.

Citations

Taiwan Centers for Disease Control. 檢驗 [Testing]. https://www.cdc.gov.tw/Category/List/fl4pGet3T9fQxrpbICzuDw

Ministry of Education. 實驗室通風換氣及生物安全櫃 [Laboratory ventilation and biological safety]. https://elearn.hrd.gov.tw/mooc/index.php&sa=D&source=docs&ust=172715516986 6138&usg=AOvVaw254SJ1lp0JJcYQWY7C3uXU