FURI | Summer 2021

Development of a Genome Editing Tool to Target Modulation Genes In Vascular Smooth Muscle Tissue

Vascular stenosis, a pathology characterized by plaque buildup in arteries, is associated with a biological hallmark: the phenotypic switch from a quiescent contractile to a proliferative synthetic phase of vascular smooth muscle cells (VSMCs). This phenotypic switch subjects VSMCs to the synthetic phase in which cell proliferation and vascular remodeling and restenosis are highly prevalent. As a consequence, arteries are narrowed and plaque rupture is initiated, ultimately causing luminal thrombosis, leading to acute and fatal syndromes. This phenotype switch is characterized by upregulation of Kruppel-like factor 4 (KLF4), a proliferative transcription factor. The objective of this project is to design and develop a CRISPR interference approach to repress the active transcription of KLF4 and attenuate the proliferative phenotype of VSMCs. In this study, the research team aims to generate recombinant adeno-associated virus particles expressing a dead Cas9 enzyme (or dCas9) fused with a transcriptional repressor and a single guide RNA (or sgRNA), directing dCas9 to target the transcriptional locus of KLF4 gene. The research team anticipates that our constructs would perturb the expression of KLF4 and exert an anti-proliferative effect on cultured VSMCs.