Part Safety
The parts we use are not on the white list. But the RUBY reporting system is essentially just a system for producing beetrophin and beetroxanthin. And this pigment is not a security threat to people and links, just a visualization system. We used promoter genes such as NCED3 and GLR2.9. We will engineer DH5-alpha,Arabidopsis thaliana Col-0 and Agrobacterium GV3101.We will order our DNA/RNA from a company, member of the IGSC.
To produce pigment, genes have been cloned in microorganisms. The main part of the development was performed on a non-pathogenic strain, E. coli DH5α and Agrobacterium GV3101.The proof of concept only was carried out on Arabidopsis thaliana Col-0.
Our plants will be genetically engineered to respond promptly to both drought and biological stresses. They will express reporter genes upon detection of these threats, causing the leaves to change color to either red or yellow, making them visually detectable by people.We will engineer DH5-alpha,Arabidopsis thaliana Col-0 and Agrobacterium GV3101.We have made new parts or substantively changed existing parts in the Registry.We will order our DNA/RNA from a company, member of the IGSC. The plants we use are standard Arabidopsis plants and have no safety concerns. The seeds of our plants are all properly preserved and do not leak into the environment.
Safe Lab Work
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.
Our Safety
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.
Environmental Safety
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.