Eric Jaffee
Research Mentor: Xin Tong
Mentor Department: Molecular & Integrative Physiology, Medicine
Author(s): Eric Jaffee, Bernardo-Ferry Abner, Xin Tong
Session: Session 1 (9:00 AM – 9:50 AM)
Presentation Type: Poster 6
Abstract
Background: Carbohydrate Response Element-Binding Protein (ChREBP) is a lipogenic transcription factor that senses sugar intake and activates genes involved in carbohydrate and fat metabolism. Our lab reported the multifaceted function and regulation of ChREBP in the liver during drug-induced hepatotoxicity and diet-induced metabolic dysfunction-associated steatohepatitis (MASH), which is a prevalent chronic liver disease among the general population with deadly outcomes, including liver failure and liver cancer. Objective: In this study, to investigate how ChREBP is involved in the pathogenesis of MASH, we will utilize Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-Cas9 Gene Editing to generate a plasmid that could be used to create a novel mouse model with hepatocyte-specific deletion of ChREBP. Methods: After designing two gRNA sequences targeting the 5’ end of the mouse ChREBP gene, we clone the gRNA sequences in the pX602 AAV plasmid, which expresses the Staphylococcus aureus Cas9 (SaCas9) enzyme under the control of hepatocyte-specific TBG promoter and drives gRNA expression under the U6 promoter. We then screen mini culture clones with restriction enzyme digestion and verify the insertion of gRNA sequences by Sanger Sequencing. Significance: This study generates a useful plasmid that has the potential to be a powerful and precise gene-editing tool to delete the ChREBP gene in vivo and uncover the molecular mechanism of how hepatic ChREBP contributes to the disease progression of MASH.
