Team:NYMU-Taipei/Safety
Overview
Our team is deeply convinced that all biological experiments carry some risk to the environment and experimenter. Under the guideline of IGEM Headquarter, we strictly follow a high standard of biosafety and responsible biological engineering. We also take note of the Laboratory biosafety manual published by WHO to ensure our lab work conducting in compliance with safety regulation. Moreover, before conducting experiments in the lab, all team members have undergone lab training by our instructors and team members in NYMU Taipei 2016. The training included laboratory emergency response procedures, operation protocols for all kinds of machines, and biological waste decontamination and disposal protocols. As one of the members of the synthetic biology community, we are responsible for abiding by the common rule.
Biosafety
Organisms We Used
We used (1) cyanobacteria Synechocystis sp. PCC 6803, (2) cyanobacteria Synechococcus sp. PCC 7942 and (3) E.coli DH5α in our project.
We extract NrtA gene from cyanobacteria Synechocystis sp. PCC 6803, which will then be transformed into E.coli DH5α. We co-culture the engineered E.coli with microalgae to induce nitrogen starvation – a pathway that will increase oil production of algae. After extracting oil from microalgae, we constructed a suicide mechanism to kill E.coli prevent contamination and potential danger of spreading engineered E.coli. To deal with the safety of E.coli, we constructed a suicide mechanism.
In pigment project, we transformed pigment genes into PCC 7942, which expressed gene and produced pigments.
Strain information of E.coli
According to the Globally Harmonized System (GHS), E.coli DH5α is not a dangerous substance. As stated in Centers for Disease Control and Prevention (CDC) Guidelines (Biosafety in Microbiological and Biomedical Laboratories, 5th Edition1), E.coli DH5α is Biological Safety Level One organism.
Strain information of Synechocystis sp. PCC 6803 and Synechococcus PCC 7942
Synechocystis sp. PCC 6803 and Synechococcus PCC 7942 are the most widely used model organisms for photosynthesis research.2,3 Both are the biosafety level one organism.
Suicide Mechanism
To prevent potential environmental danger, we created a suicide mechanism to kill E.coli. We designed suicide genes, endolysin and holin, to clear away all of engineered E.coli, ensuring the safety of E. coli. Endolysin and holin are similar to the mechanism used by team Pecking (2014 iGEM Beijing). Holin can induce holes on cell membrane, allowing Endolysin to pass through the membrane and decompose peptidoglycan. After the cell membrane and cell wall are destroyed, E.coli will be lysed.
Lab safety
All experiments are done in Biosafety Level 1 (BSL1) laboratory. Our lab is equipped with safety facilities, such as emergency shower and fire extinguishers, and we conduct experiments under Laboratory Safe Hygiene Precautions of our school.4 In addition, there are safety insert and postcards all around the lab as reminders. Lastly, all experimental protocols were checked and guided by our PI, Dr. Chou before we conducted.
Lab training
Every members of wet lab in NYMU iGEM team have finished lab safety lesson and training twice, which are directed by professors who instruct us step by step. Thus, members can conduct experiments properly and safely.
Protection
In our security-controlled laboratory, each member wears laboratory coat and gloves with long pants and shoes of full coverage. When we leave the lab, we make sure to remove our coat/gloves and wash our hands with antibacterial hand wash. We strictly ensured that all of these are done in addition to avoid touching door knobs with gloves to prevent carrying bacteria beyond the lab.
Waste
We gather chemical and bacteria mixture into a bottle containing bleach after conducting the experiments. Besides, we sterilize all trash with autoclave and put it at a specific area of contaminative trash. All the wastes in our lab are handled seriously and strictly to prevent contamination.
Reference
- U.S. Department of Health and Human Services. (2009). Biosafety in Microbiological and Biomedical Laboratories (5th ed.). HHS Publication No. (CDC) 21-1112
- Bertram M. Berla, Rajib Saha, Cheryl M Immethun, Costas D. Maranas, Tae Seok Moon, and Himadri B. Pakrasi. (2013). Synthetic biology of cyanobacteria: unique challenges and opportunities. Front Microbiol, 13(4), 246.
- Raul Muñoz, Cristina Gonzalez-Fernandez. (2017). Microalgae-Based Biofuels and Bioproducts: From Feedstock Cultivation to End-Products. Woodhead Publishing.
- Taiwan National Yang Ming University. (2010). Laboratory Safe Hygiene Precautions. (http://ces.web.ym.edu.tw/ezfiles/151/1151/img/183/laboratorysafehygieneprecautions-991001.pdf)
