THC Sensing, reporting and metabolizing system

This year, borrowing ideas from the modified Pmr system of 2021 BNU-China and the anti-THC antibody of 2019 Queens_Canada, we replaced the iron (III) binding motif with the anti-THC antibody which enables the specificallydownstream genes expression to Δ9-THC. We created the new Δ9-THC sensing mechanism and added it into the prokaryotic system, providing a basis for the subsequent synthetic biology design that may targets at Δ9-THC.

At the same time, it has been a difficult problem to recieve the information conveyed by intestinal bacteria. But in our project, when sensing Δ9-THC, engineered bacteria express and secrete C-phycocyanin, thus changing the color of feces. We use this unique idea to report the pathway induced by Δ9-THC, and provide a new visual test method, achieving regular monitoring. Our reporting system provides a new prospect for the development of reporter synthetic biology.

It’s brilliant that we successfully constructed the prokaryotic systems to surface display CYP2C9, CYP2C19 and UGT1A3, which contributes to the metabolism of Δ9-THC and the prokaryotic manufature of these enzymes.

Useless killer

We verified through a series of experiments that the mazE and mazF systems under the action of temperature-controlled RBS have little effect in inducing death of engineered bacteria, but we cannot exclude that it may be the effect of laboratory conditions and experimental manipulation factors. We hope that subsequent iGEM teams will use them with caution.

handbook

In the early stage we encountered a lot of problems that arise for new teams. We believe that many problems are common, so we recorded many basic problems encountered in the process of team formation and competition, and gave our solutions to compile this experience manual to provide reference and help for later teams, effectively stimulating the interest and even love of synthetic biology among students around us, lowering the threshold for them to participate in the competition, and promoting their understanding of iGEM events.

Many thanks to JLU_China for their invitation to us to participate in this handbook compilation. We sincerely hope to contribute and support the subsequent participants of iGEM competition!!!