Team:CSMU Taiwan/Safety

CSMU - iGem



















Safety






General concern and regulation of biosafety

Overview

iGEM2020 CSMU_Taiwan team is in full compliance with the iGEM's lab safety rules and security policies of the iGEM competition. All experiments of our iGEM project were conducted in the Biosafety Level-1 laboratories under the regulation of Environmental Safety and Health Center of Chung Shan Medical University. Team members in CSMU_Taiwan were all well-trained by our mentors and instructors and had the ability to conduct experiments safely. Our advisors also taught us how to use the instruments and chemicals in our laboratory safely.

Lab Safety

People who entered the laboratory should follow the established laboratory safety principles of our school. People should wash their hands upon entry into and go out of the laboratory. When doing experiments, wearing suitable clothing with gloves and lab coats is necessary. To the waste of biochemical experiments, all kinds of consumables in the working area of the laboratory are considered biohazard waste. Those biohazard wastes would be collected in a specially labelled red plastic bag and sent to the Environmental Safety and Health Center of Chung Shan Medical University for the further disposing process.

Project Safety

Our experiment

This year, our main experiments were conducted in vitro. The only species we use in our experiments is E. coli DH-5 alpha1, which is the chassis in our project. The genes we used contain two main parts, toehold switches and the genes of expressed proteins, which were both synthesized directly from biotech companies. We designed toehold switches ourselves based on three papers2,3,4. They can combine with different miRNA and then open their stem-loops. We chose GFP and invertase as our expression proteins. Both of toehold switches and expression proteins are not harmful to human beings and creatures. Our project would be completely safe under the situation of operating correctly.

Final project

Our final product is a detector of oral cancer. We design our kit, miRNA.doc, based on glucometer. The new kit can work safely as the original one who has passed the standards of medical equipment. One of our primary concern is the entry-level for using miRNA.doc. We attempted to simplify the process as much as we could. However, the use of chemicals is unavoidable and would require basic chemical safety knowledge to use the kit.

Reference

  1. Bryant FR. Construction of a recombinase-deficient mutant recA protein that retains single-stranded DNA-dependent ATPase activity. J Biol Chem. 1988 Jun 25;263(18):8716-23. PMID: 2967815.
  2. Green, A. A., Silver, P. A., Collins, J. J., & Yin, P. (2014). Toehold switches: de-novo-designed regulators of gene expression. Cell, 159(4), 925-939.
  3. Pardee, K., Green, A. A., Takahashi, M. K., Braff, D., Lambert, G., Lee, J. W., ... & Daringer, N. M. (2016). Rapid, low-cost detection of Zika virus using programmable biomolecular components. Cell, 165(5), 1255-1266.
  4. Wang, S., Emery, N. J., & Liu, A. P. (2019). A novel synthetic toehold switch for microRNA detection in mammalian cells. ACS synthetic biology, 8(5), 1079-1088.