Team:Yonsei Korea/Safety
In the Laboratory:
The Science
Behind It All
To help our team initially with designing and conducting experiments in a safe and secure manner, all research members completed lab safety feature tours (or already completed them from prior internship or lab research experiences). Although we initially had all research members in the lab at rotated times to try experiments, we shifted to having only research members with prior or current lab experiences work in the lab. This was beneficial additionally in part as experiments were conducted more efficiently and safely as our experienced members had all the knowledge for the lab work.
We were not only concerned with the safety and security of experiments such as risks of leakages, contamination, burns, etc. Unfortunately, in South Korea the daily number of coronavirus cases has been on the rise and many social distancing regulations have been put in place. As a result, we limited the number of members working in the lab at a time to 2-3 members in order to maintain a safer working environment for ourselves as well as the other researchers and students in our lab. In part this worked in line with maintaining proper distancing and open space not to spread the virus, but also enabled our members to work more efficiently and closely with the experiments, thus reducing greater risks of misconduct or accidents.
According to the Ministry of Food and Agriculture in South Korea, carrying out experiments with the pathogen Magnaporthe oryzae is strictly prohibited unless in designated laboratories. Hence, we did not use any organisms directly from this pathogen in our lab. We performed cell-free experiments using only DNA sequences synthesized by companies such as BIONICS and IDT. Our lab did not use any animals or living organisms as this comes with added risks and safety concerns inside the lab as well as to the environment. Nevertheless, we still procured and employed certain hazardous chemicals such as heavy metals, carcinogens, acids, and corrosive chemicals. For example, for agarose gel electrophoresis: 5x TAE buffer, for PAGE experiment: acrylamide gel 40%, APS 10% (Ammonium Persulfate), TEMED(Tetramethylethylenediamine), 5x TBE (Tris Boric-EDTA) buffer, for F-8 DNAzyme reaction buffer: CuCl2, MnCl2, MgCL2 (all in powder form), for nanoparticle-based experiments: DTT (Dithiothreitol) solution. In these cases, all members handling these chemicals wore safety goggles, a fitted lab coat, and gloves. After all experiments were conducted, all used benches and materials were cleaned and stored appropriately. At the end of each lab day, members were prompted to wash their hands thoroughly to avoid carrying any contaminations from the lab nor spreading the coronavirus. On the same note, all members wore masks in the lab all the time as this was the regulation for our research institution as well as our personal responsibility.
As aforementioned, doing experiments involving Magnaporthe oryzae is prohibited in the majority of laboratories in South Korea due to its risk as a bioweapon noted by the Korean Ministry of Food and Agriculture. Accordingly, we did not do any DNA extractions of the pathogen. We had 100 base pair long regions of the target mif23 gene of our pathogen synthesized by a provider (BIONICS/IDT). Also, we designed and synthesised an F-8 deoxyribozyme that specifically cuts the target sequence. Using these synthetic DNA oligonucleotides, we performed a deoxyribozyme catalyzed cleavage reaction in a reaction buffer prepared according to the protocols presented in research articles. We used the salt-aging method to functionalize gold nanoparticles with thiolated DNA that is complementary to the DNA products cleaved from our target mif23 gene.
Apart from these safety and security measures, ethics were also considered throughout many stages of our project. Ensuring the safety of our team members and environment is one of our main priorities; nonetheless, ensuring we carry out all our responsibilities and endeavors responsibly and ethically was also one of our key practices.
Throughout our efforts in our human practices, we were able to travel to a rice farm in Korea and gain many valuable insights from the farmers there. We made sure to hear their perspective on their needs and pains when it comes to pathogens affecting their crops as well as explain our concept and how it might help them. In addition to this, we also considered the future ethics that could stem from the further development of our research. In order to reduce any risks of harmful chemicals or organisms released into the environment, our concept emphasizes simplicity. A tool or device that can be used anywhere and by anyone without the need for special equipment or harmful chemicals.
Yonsei_Korea is a convergent team of students based in Seoul, South Korea striving to create meaningful change utilizing synthtic biology in the iGEM 2021 Competition.