Ronak Arun
Pronouns: he/him
Research Mentor(s): Adelaide Tovar
Research Mentor School/College/Department: Computational Medicine & Bioinformatics / Medicine
Program: UROPF
Session: Session 7 (4:40pm – 5:30pm)
Authors: Ronak Arun, Adelaide Tovar, Jacob Kitzman, Stephen, Parker
Abstract
Diabetes is a common illness caused by genetic and environmental factors that plagues many people worldwide. Though specific genetic associations with diabetes have been identified, few of these have been translated to new drugs and treatments. This project focuses on genes associated with type 2 diabetes (T2D) that are also known to cause rare forms of diabetes known as MODY (Mature Onset Diabetes of the Young), a genetic condition that mimics the symptoms of T2D. For this project, we are using CRISPR-Cas9 to knock out NKX6-1 and HNF1B in the EndoC-ßH1 and -ßH3 human beta cell lines. Eventually we will perform replacement with mutated forms of each protein to observe the phenotypic effects associated with certain allelic variants. To knock out our genes of interest, we will infect these cells with a virus carrying Cas9 protein along with specific guide RNA (gRNA) sequences. We will perform DNA genotyping, gene expression analysis via qPCR, and protein expression analysis via Western blots to determine whether the knockouts were successful. In other studies about genetic conditions, this project can serve as a guide for how to study the effects of transcription factor mutations on gene expression and disease processes. More specifically, this project will give us information about the specific interactions between mutated forms of NKX6-1 and HNF1B on key diabetes-associated genetic pathways. Hopefully this information can be used in the future to provide clinical predictions for people who carry mutations in these genes.
