Prashant Bhat Rose Hills
Investigating the Mechanism of RNA-Mediated Target Interference in a Bacterial CRISPR System
CRISPR RNAs (crRNAs) are instrumental in mediating a prokaryotic immune response by targeting and subsequently directing the degradation of invasive genetic elements. Research is currently underway to harness the CRISPR system for biomedical applications including the detection and diagnosis of viruses in human clinical samples. My project investigates the components of the large multi-subunit complex found in E. colitermed Cascadewhich is responsible for targeting and destroying invading nucleic acid via an unknown mechanism. Prior work from our lab has shown that a key protein in the Cascade complex called Cas7 may be essential in unwinding the invading dsDNA duplex, thereby making it accessible for degradation. Using a combination of biochemical tools and x-ray crystallography, I plan to investigate the structural interactions of Cas7 with a DNA: RNA hybrid to examine the method of target recognition by the Cascade complex, with the ultimate goal of an atomic resolution structure of this critical Cascade subunit bound to target RNA.