FURI | Fall 2021

Development of CRISPR Interference Constructs to Modulate the Phenotypic Switch of Vascular Smooth Muscle Cells

Previous studies highlight the critical role that upregulation of Kruppel-like factor 4 (KLF4) plays in promoting the pathological phenotype of vascular smooth muscle cells (VSMCs). This FURI project adopted a CRISPR Interference (CRISPRi) approach to perturb KLF4 expression and exert an anti-proliferative effect on cultured VSMCs. Specifically, small guide RNAs (sgRNA) targeting the KLF4 gene locus were subcloned to a viral vector carrying a nuclease dead Cas9 fused with a transcriptional repressor. The research team is producing recombinant viral vectors (rAAV) and will further determine the extent to which the CRISPRi constructs suppress KLF4 expression and the pathological phenotype of VSMCs.