"Development and Characterization of DNA-targeting enzymes against HBV"
Chronic hepatis B virus (HBV) infection is the most common risk factor for liver cancer worldwide. This project aims to develop an alternative method for treating HBV infection by utilizing the areas of expertise contained in two labs at the Hutchinson Center.
We are currently working to develop HBV-specific DNA-targeting enzymes (homing endonucleases (HE), zinc-finger nucleases (ZFN), and transcription activator-like effector nucleases (TALEN)), which can specifically recognize sequences in HBV DNA and induce double-strand DNA breaks. The repair mechanism for these breaks is error prone resulting in inactivation of genes near the cleaved target sequence, which in turn eliminates the replicative capacity of the virus. The goal of this proposal is to facilitate this work by comining the biophysical, structural and enzymological capabilities of the Stoddard Lab with molecular biology and virology studies in the Jerome Lab. From Dr. Barry Stodard, expert in structural and biophysical enzymology, I plan to acquire experience in analysis and characterization of candidate enzymes in order to ptimize their antiviral activity (Aim 1). Similarly, with the assistance of Dr. Keith Jerome, expert in laboratory models of latent viral infections, I will design and perform experiments to test DNA cleavage and mutagenic activity in both a reporter cell system and physiologic model of HBV infected cells (Aim 2).
The interdisciplinary nature of this project and the complementary expertise of the two labs will greatly broaden the scope of my postdoctoral training and should allow this important project to move forward as rapidly as possible.