Team:Sorbonne U Paris/Safety
Safety
For our project we built a genetically modified organism so we had to be responsible about our biological engineering. Additionally, working in a lab involves security rules depending on the kind of reagents, machines and techniques we use. This year being special because of the pandemic we are very grateful to have had the opportunity to work in a lab, but this meant to adapt and add more security rules.
Safe project:
The first concern to tackle is to work with a safe chassis. Indeed, the organism we used for our project is the microalga Chlamydomonas reinhardtii strain D66. This is a non-pathogenic strain classified as a GRAS organism, so there were no risks for us nor our colleagues working in the same laboratory. Then, we worked safely and securely with our parts.
The actual knowledge on Genetically Modified Organism (GMOs) cannot assure us of the consequences GMOs have on our environment and health. Thus, we have to be extremely careful when using them and consider the risk of contamination. In our case we want to build a filter of GMO algaes implemented in a water filtration system. In order to prevent the spread of our alga in the natural environment, we developed a kill-switch system for Chlamydomonas reinhardtii. By modifying the alga we make it dependent on a specific condition (in our case the presence or absence of ultraviolet light) to survive. We made our alga intolerant to UV radiation so that when it is inside a bioreactor opaque to UV rays, these genetically modified strains would live normally and be able to purify the water from harmful compounds. However, if they accidentally escaped the tank they would die because exposure to UV light leads to the activation of their dying process. For this method we were inspired by the iGEM team TU-Munich 2013. Their “kill switch” relied on making the survival of their transgenic moss dependent on a specific infrared wavelength.
Safe lab work:
During the iGEM competition, all experiments were conducted in the Laboratory of Computational and Quantitative Biology (LCQB) depending on Sorbonne University and CNRS (French National Centre for Scientific Research) under the surveillance of researchers (Assistant professor or PhD student) who were already very familiar with the experimental procedures and practices we were using. Therefore, they showed us and explained every safety and security measures prior to bench work. First of all, we always wore cotton lab coats and appropriate clothing (long pants, closed shoes and hair tied back) when in the lab. We always wore protective gloves and changed them regularly to prevent contamination of our samples. In the EU, the CLP regulation (classification, labelling and packaging of substances and mixtures) or the Regulation N. 1272/2008 informs users about the risks of hazardous chemicals through their labelling with standard symbols and the creation of safety data sheets in order to protect human health and the environment. Most of the components used in our experiments are classified as safe. However, for our project we prepared solutions of pollutants to be tested on the
Chlamydomonas reinhardtii
microalgae. All were done very carefully under a chemical hood. We also used Ethidium Bromide during our experiments. It is an intercalating agent used as a fluorescent dye to visualize DNA in gel electrophoresis. According to the EU Regulation N. 1272/2008, Ethidium Bromide is acute toxic, irritant and a health hazard. Moreover, it is mutagen and probably carcinogen. For this purpose, we always wore protective gloves while handling this substance in a delimited area. When manipulating Ethidium Bromide, we were extremely careful not to touch and thereby contaminate other surfaces outside the area and changed gloves before going to another area. All the tips, gloves and other disposable objects contaminated with Ethidium Bromide were disposed of in a specific trash bin.
After the experiments, we dropped the contaminated waste in specific bins and we always disinfected work areas and washed our hands with soap before leaving the lab. Finally, we can find in the laboratory common safety equipment like eye wash stations, showers, fire extinguishers and emergency cabinets that can be used in case of an incident.
COVID-19:
Due to the pandemic situation we had to sign authorisations for each member that had to work in the laboratory. A limited number of people were allowed to be present in the lab at the same time, 1 meter physical distanciation had to be respected and we had to never be alone for obvious safety reasons.
We also had to wear a mask at all times including during the manipulations and wash our hands when arriving and leaving the lab. Additionally, we had one lab coat per person and we were not allowed to share it, likewise for the protection glasses.
References:
- Henkler F, Tralau T, Tentschert J, Kneuer C, Haase A, Platzek T, Luch A, Götz ME. Risk assessment of nanomaterials in cosmetics: a European union perspective. Arch Toxicol. (2012).
- Homodium Bromide : Safety and Hazards. PubChem. Available on pubchem.
- Murbach TS, Glávits R, Endres JR, Hirka G, Vértesi A, Béres E, Szakonyiné IP. A Toxicological Evaluation of Chlamydomonas reinhardtii, a Green Algae. Int J Toxicol. (2018).