This year, NUS_Singapore-Sci has sought to find out the myths and misconceptions about gene editing (#CasAsks) among the public, to identify which aspect of gene editing we should focus on for our project. Through surveys and interviews with various stakeholders, we identified that the main concerns were the permanent nature of DNA editing and ethical considerations. This made us decide to focus on RNA editing as our project. We have also attempted to debunk the myths and misconceptions via school outreaches (#CasTeaches) and educational videos (#CasTalks) after speaking to scientists and people in the field. With all the knowledge on hand, we have summarized and compiled all the interviews into #CasWrites. You can hop over to our website to find out how all of these are integrated into our project!

We sought to improve BBa_K2083010, a GFP with a mutated start codon from ATG to ACG through our own dual color reporter system BBa_K2807013 which is made up of mCherry and eGFP connected by a T2A linker containing a mutated start codon of ACG. It not only allows a robust OFF-TO-ON switch from the ACG mutant to wild type (ATG), but also demonstrates strong linear correlation between EGFP and mCherry fluorescence, allowing normalization of transfection and expression levels. More information can be found here, detailing on how the improvements were made.

We have attempted three types of modelling for our project which involves modelling a suitable linker for the RESCUE Editor, hidden concentrations of the RESCUE dynamic system and lastly, and estimating the parameters for the RESCUE system. Head over to our website to how the modelling was done and how it was integrated into our project.

The RESCUE Reporter system was tested in HEK293T mammalian cell lines transfected with base editor 3 and 4 made up of rAPOBEC1 and dCas9. It could successfully report DNA editing events under realistic conditions. Take a look at our results for more information on the demonstration.