Examining the function of Notch1 and Dach1 on collateral artery formation in the mouse brain through CRISPR-mediated inhibition and activation.

The number one cause of death is coronary artery disease (CAD), which results from narrowing and occlusion of coronary arteries.  A potential treatment could be induction of coronary collateral arteries to bypass blocked vessels.  Collateral arteries in humans are associated with increased survival from CAD, but there is a lack of knowledge surrounding their formation. Two genes that have already been identified as important in artery development are Notch1, a key cell fate determinant, and Dach1, which has been shown to increase collateral artery formation in the heart when overexpressed (Raftrey et al. 2021).  These two genes will be tested in the context of collateral artery formation, i.e. the postnatal mouse brain.  These two genes will be inhibited with inducible CRISPR-mediated inhibition or activated with inducible CRISPR-mediated activation, with adeno-associated viral vectors (AAVs) used to deliver guide RNA for target genes.  Evaluating these genes has aided in optimizing our procedure so that future experiments can test multiple genes of interest at once.  Identifying the molecular mechanisms of collateral artery formation may aid in the development of protective treatments for humans against ischemic diseases.