Team:NTNU-Trondheim/Safety

SulFind 2021

Safety

Overview

Early on in our iGEM cycle, we began the process of ensuring adequate safety regarding our project design and our own safety when working in the lab. Eventually we ended up with a design that does not contain any living organisms, thus dramatically reducing the risk of escaping and interference with the environment. Before finalizing the project design and moving away from living organisms, we used E. Coli strains that are in risk group 1 to not endanger humans and the environment unnecessarily.


Safety on Project Design

As one of our two candidate designs consist of modified E. coli cells, there is always the risk of the modified bacteria escaping the lab and interfering with the environment. Therefore we very carefully followed all regulations regarding work with recombinant DNA and GMO whilst working with the design involving living cells. However, our final design utilizes non-living heme proteins. This reduces risks tremendously as compared to using living organisms in the design. Proteins are easier to contain in the lab, as they are unable to replicate and spread in the environment like bacteria. By choosing protein instead of bacteria, we provide ourselves with more control.

Furthermore, when working with living cells we chose E. coli strains DH5a, and BL21, which are in risk group 1. This way, risks concerning the host organism are reduced. For this design we also chose GFP which emits fluorescence and hence is a non-harmful reporter gene. The probe we used in our final design, Cyanine3 NHS ester, also emits fluorescence and is a non-hazardous chemical.

Our final design will be embedded on a microfluidic chip. This produces a strictly controlled system, where we accurately control what is passing in and out of the tubes on the chip. It allows us to directly and safely collect the waste leaving the chip and dispose of it in a correct and secure manner. In addition, the use of a lab-on-a-chip means working with concentrations on nano scale. This reduces risks related to project design, because if an accident were to occur, the amount spilled would be very little compared to having a project design on macro scale.


Safety in the Lab

As we were lucky enough to execute actual lab work this year, we wanted to make sure that all necessary safety precautions were put into place when it comes to our lab work. First of all, we all underwent safety training before gaining access to the Molecular Genetics and Microbiology (MolGen) lab at our university. Since this lab is certified for risk 1 and 2, there are several obligatory activities you must do first. These activities consist of an online lab course followed by an online lab exam, and a guided tour in the lab where you must perform several activities like sorting lab equipment before their waste disposal. After our initial safety training, we got thorough tutorials on lab equipment by trained personnel in the lab, to ensure that we used equipment properly.

As stated in the MolGen lab guidelines, we wore PPE (personal protective equipment) at all times and had extra safety measures when working with hydrogen sulfide. None of us ever worked alone, and when dealing with (H2S) we worked under a fume hood. All hazardous waste was collected and disposed of according to university guidelines.

To ensure that no accidents happened at the lab, we made thorough lab protocols for all of our experiments. These protocols can be found here. Lastly, we did a detailed risk assessment for our lab’s supervisor.


Guidelines

Norway

In Norway, all research practices regarding genetically modified organisms are regulated by the Gene Technology law [1].


University

All laboratory activities occurring within the Norwegian University of Science and Technology iGEM Team are regulated by the university’s HSE Division [2].


General lab rules for IBT laboratories

Lab rules for the general microbial molecular biology laboratories [3].


References

  1. Lovdata. Lov om framstilling og bruk av genmodifiserte organismer [Internet]. Oslo: Lovdata; 24.08.2021. Available from: https://lovdata.no/dokument/NL/lov/1993-04-02-38
  2. NTNU. HSE guidelines - summary pages [Internet]. Trondheim: NTNU; 28.10.2013 [updated 18.08.20]. Available from: https://innsida.ntnu.no/wiki/-/wiki/English/HSE++guidelines+-+summary+page
  3. NTNU. HSE at IBT [Internet]. Trondheim: NTNU; 14.01.2019 [updated 02.08.21]. Available from: https://innsida.ntnu.no/wiki/-/wiki/Norsk/HSE+at+IB