Home Team Team Attributions Collaborations Project Description Design Proof of concept Engineering Results Notebook Implementation Contribution Experiments Parts Safety Human Human Practices Communication Partnership Jamboree Organization Awards Education Model Sustainable Safety Safety Safety is a really important part in the iGEM competition as well as in daily labwork. In this part of the wiki, we, iGEM Unizar, want to explain how we have improved our project to take it to another level of security, not only in labwork but also in ethics. Microorganisms Different Escherichia coli strains have been used in our project. For cloning purposes E. coli XL1-Blue E. coli DH5α For nanobody expression E. coli UT5600 The use of E. coli was chosen due to its lack of pathogenicity and ease of cultivation. All the strains ara cataloged as level 1 of security by the Spanish legislation. Lab workspaces These organisms were manipulated not only in open-bench workspace but also in biosafety cabinets for providing sterility. Therefore, in our project we have combined the usage of level 1 and level 2 workspace. For the correct usage of a level 2 workspace, the team was trained in a biosafety and security course by Carmen Lafoz, technician of the Microbiological Analysis Service and Member of the Biosafety Committee of the University. Working in lab While working in the lab we wore IPE (individual protection equipment) according to the risk. Basic IPE in our lab is composed by: Gloves for manipulating micoorganisms and DNA. It is well known that DNases are everywhere, so the usage of gloves can minimize the risk of DNA degradation. Gloves also protect our hands from chemical reagents. Lab Coat for avoiding different splashing. Surgical mask due to the COVID-19 pandemic restrictions. Working in the lab is regulated by RD2009-27" Spanish legislation. Therefore, we made sure that we were following Good Laboratory Practices. Security and safety protocols at the lab In our institution (University of Zaragoza), there is an Occupational risk prevention unit, which is in charge of making protocols to be followed at the lab and ensuring security. This document has all the guidelines about prevention and use of different materials at the lab. For example, the use of Bunsen burner must be with out gloves preventing burns Security lab manual. This document has all the guidelines about how to manage the biological waste at the lab. Waste managment. Luckily, we had no accidents, but if there had been any we should have followed this manual. In case of emergency. Risk in our project Biological and chemical risks were present in our project. As, we identified them, we could improve safety in our expriments. Example of improved security Chemical Risk→Our project is based on nanobody expression in bacteria, for which a plasmid is needed. We use for plasmid purification some kits such as ThermoScientific. This kit has sodium hydroxide, a corrosive chemical, so the person in charge of purification wore gloves, a coat, and security glasses. After purifying everyone washed their hands. Biological risk→ Different CDRs were recombined in order to create new nanobodies, so we used synthesized DNA. The person who manipulated them always wore IPE and at the end everything was disinfected with ethanol 90%. Even though E. coli is not a dangerous pathogen for human health, it is important to work with it in a cero risk way. To do so agar plates we disposed of in a special bin for biological residues and liquid cultures were neutralized with bleach. GMO and RECONBY GMO (genetically modified organisms) are defined as any organism modified by any genetic engineering technique. There is strong legislation (RD452/2019) about constructing them due to ethical purposes. We haven't planned to release them to the environment complying with the iGEM policies. Nevertheless, if they were released, they wouldn´t survive. Laboratory strains like this one, without optimal conditions are incapable of living in the environment. Furthermore, if our purified nanobodies ended up in the environment, there would be no risk of dissemination since they are not living parts capable of replicating. Safety lecture Appart from protocols, rules, EPIs and other security items, we wanted to know deeper. This is the reason why we took a lecture with Carmen Lafoz Pueyo. She is laboratory technician at the University of Zaragoza and a laboratory safety specialist.
