Genomics Approaches to Investigating HCV: A Molecular Recorder and CRISPRi Screen

Abstract

Chronic hepatitis C virus (HCV) infections often lead to significant long-term complications, including permanent liver damage and hepatocellular carcinoma. Currently, the molecular mechanisms driving cancer development in patients with chronic HCV are unclear. To untangle the contributions of direct effects on infected cells and bystander effects on cells near the infection site, I developed components of what will be an inducible molecular recorder for interferon signaling in human hepatocytes. By designing, cloning, and testing a Cas12a effector and guide RNAs, as well as cloning and evaluating several liver cell lines containing the necessary genome editing machinery, I identified a highly efficient guide RNA to be used in the final recorder system. This larger project led by Dr. Aaron Lin will ultimately allow for greater insights into the molecular mechanisms underlying the development of hepatocellular carcinoma from chronic HCV infections.