Our Original Idea - PlaBreze
          The Lethbridge High School iGEM Team starts the year off with a brainstorming session to determine what our yearly project will be. Our first choice was not “Dia-Beatable”, but a different project dubbed “PlaBreze.” The prototype name was a goof of the existing air refresher company “Febreze.” However, due to copyright concerns our team never intended to keep the name and change it later down the line. Though, as Plabreze would be scrapped which will be further explained below, the name never reached such a stage.

          PlaBreze was pitched by members Steven Yang and Morteza Faraji. The project was focused on genetically modifying houseplants to emit non-natural scents such as cotton candy, processed chocolate, processed vanilla, etc. Below is a snapshot of the original pitch presentation from Yang and Faraji.



          The purpose behind PlaBreze was to improve people's mental health through comforting scents. The team noted the unsustainable and expensive costs of current air refreshers and PlaBreze was intended to be a more environmentally-friendly and cheap alternative. Furthermore, the team believed that PlaBreze's plant-based medium would increase global oxygen supply through the plants' production of oxygen.

          Our team even had a prototype prepared using the Pothos Ivy houseplant and the Vanilla Synthase gene. Additionally, former team member, Alexandru Albu created the following logo for PlaBreze based on Febreze's logo:



          However, PlaBreze was scrapped after the team presented the project to the judges during MindFuel's Tech Futures Challenge. Judges pointed out that many consumers would prefer buying natural plants over PlaBreze-produced plants, and that there were more efficient means of improving mental health, rendering PlaBreze useless. They were also concerned with the potential for PlaBrese plants to become invasive. The team realized that our iGEM project needed to have a bigger global impact than mental health through nice-smelling plants. Thus, PlaBreze was scrapped.

          Unfortunately, the team had greatly enjoyed working on PlaBreze since the entire time pitched in to create a promotion video and presentation for MindFuel. Thus, when PlaBreze was scrapped, our team grew demoralized since we had to return to the pitching process. Though, we ensured that every pitch matched the iGEM criteria and had the needed global impact so we wouldn't need to scrap another project. Finally, we turned our focus to our new project… “Sugar Baby!” 

          “Sugar Baby” was the original name for “Dia-Beatable.” The project itself was pitched by members Sophia Bird and Aubrey Nilsson. However, the team later realized that the name's had a possible association with inappropriate connotations, so the team was forced to brainstorm a new name for the project. Various names were suggested in this period, including “Let Them Eat Cake!” a nod towards Marie Antoinette, and numerous (far, FAR, too many to name) S.U.G.A.R acronyms. But, it was Aubrey's suggestion of “Dia-Beatable” that won the team polling to become the project's official name.


RNA Aptamer Design
          An important feature of our project will be the ability to control translation of insulin in response to a patient's needs. We don't want to continuously produce insulin if it is not needed as it wastes resources and can lead to extremely low blood sugar levels. Therefore, we chose to design an RNA apatmer that would be used as an "on/off" switch for insulin translation. Our first design proposed using a glucose aptamer to control downstream translation of the insulin gene. When glucose is not present, the RNA would form a structure that would occlude the ribosome binding site (RBS), thus not allowing translation of the downstream gene. Upon binding of glucose, a conformational change within the RNA would be triggered and the RBS would become available for binding and thus translation of insulin in response to high levels of glucose could occur.





          We decided to test the ability of our proposed glucose aptamers in vitro by using an RNA Mango biosensor. We hypothesized that binding of glucose by the glucose aptamer would cause an increase in correctly folded RNA Mango, thus resulting in higher fluorescent output. After our experiments (Proof of Concept), we realized we would need to refine our aptamer sequence.





          Following the BioTreks conference and feedback from judges, we also found that having a glucose "on/off" switch would not be the best way to control insulin translation levels. Since transcription and translation will happen within cells and the glucose that we would like to measure is in the blood, our biosensor would not work as we had hoped. Therefore, we chose to design an insulin aptamer instead. This RNA structure would be able to bind to the insulin protein and prevent any translation of the downstream insulin gene. As the cellular level of insulin decreases, the insulin protein would dissociate from the insulin aptamer allowing for ribosome binding and translation to occur. This new design would be better able to control the level of insulin needed.