All our work is covered by the White List.
FIAT LUX is based on a metabolic reaction of intracellular substrates to produce luminescence. The products of the reactions are recycled thanks to other metabolic reactions, so bacteria do not produce extracellular products or molecules: there is no risk of creation of hazardous substances (see Description page for more information).
Table 1 - Risks associated to the genes used in the wet lab
To produce bioluminescence, genes have been cloned in microorganisms. The main part of the development was performed on a non-pathogenic strain, E. coli DH5α. The proof of concept only was carried out on Dickeya solani, a phytopathogen.
Table 2 - Risks associated to the bacterial strains used in the wet lab
Although our project involves helping farmers and agriculture, FIAT LUX is not intended for use in fields or in plants in nature. Indeed, it is not a treatment made to be used by the farmers themselves, but a research tool that will be used by scientists in a biosafe and closed laboratory. For example, depending on the bacteria they are handling, scientists may be required to work in a level 3 biological safety laboratory. In this type of biosafe laboratory, safety rules ensure isolation and total containment of infectious agents within the laboratory. Thus, the risk of accidental release of genetically modified microorganisms by ilux in the environment is drastically minimized.
Safety concerns are mainly regarding the bacteria we are manipulating. The genetic modifications we introduced do not increase their dangerousness. That is why we have worked in academic laboratories classified level 1 of biosafety (standard microbiological lab).
Safety training was received by the team before starting any experiments, and a safety officer was always available in the lab for any questions or doubts we may have. We also participated in a safety workshop hosted by iGEM. Before the experiments, all our protocols were verified by our experienced professors or advisors, and supervision by them or any researcher of the lab was always ensured. We also relied on Florence Ruaudel, an institutional biosafety officer at the laboratory we worked in.
Appropriate and required protection was used during all steps of wet lab manipulations.
Among protocols that required a special safety consideration:
- A particular attention was taken to the handling of EtBr for electrophoresis analysis. It was manipulated on a special bench. Gloves were changed after any handling of the product, and any tool that touched this bench stayed on the bench, which also comported a special water tank to wash the material.
- FFP2 masks were worn for manipulation of concentrated antibiotics, and the manipulations were done under fume hood.
- When using a flame for sterility, no inflammable material was approached.
- When using UV light, appropriate protection glasses were worn.
- Acids and corrosive chemicals were handled with care and thrown out in the appropriate chemical container.
About the laboratory arrangement, there is one room by manipulation type, to isolate the manipulation of dangerous substances, and not transfer dangerous substances from one room to another. For example, we have an electrophoresis room, a media preparation room and microbiology rooms.
We also used a specialist greenhouse.
In our laboratory, when we manipulate genetically modified or pathogenic microorganisms, biohazard containers are used. Two types of sharp safe boxes are used: plastic bags for non-stick biological waste and rigid boxes for sharp waste. Tools and reusable lab materials are autoclaved after each use. A specific area is dedicated to “dirty” material, separated from the “clean” area, to prevent contamination.
Everything concerning the infection of plants and media preparations will be completed in a level 2 lab space (BSL-2), ensuring that the risk of accidental release of our GMO is minimum. The infected plants were eliminated with the infectious waste.
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