Poster #036
Comparing Minimal Promoter Activity for CRISPRa Reporters
Mentors: Nik Lind, PhD; PI: Julian Halmai, PhD
CRISPR activation (CRISPRa) is a powerful epigenetic tool that upregulates gene expression without modifying the underlying DNA sequence, enabling treatments of genetic diseases. One significant challenge is our ability to reliably predict a gene’s responsiveness to epigenetic modification based on chromatin context. To address this, we developed a GFP-reporter gene to analyze CRISPR-based epigenome editing at various chromatin environments. To identify a promoter sequence with low basal expression, we compared the efficiency of an 87-bp super core promoter (SCP) and a curated 20-bp YB_TATA promoter to drive GFP expression in HEK293 cells. Additionally, we tested the reporter’s ability to be upregulated by CRISPRa by targeting each promoter with dCas9-VP64. This approach allows us to determine gene responsiveness without confounding sgRNA sequence variability. These results will contribute to our understanding of how chromatin context influences epigenome activation, permitting further development of CRISPR-based gene therapy through CRISPR strategies.