Addressing Barriers to Gene Therapy: Safety and Efficacy

Abstract

The development of the CRISPR/Cas9 gene editing system has driven renewed interest in the field of gene therapy. However, despite recent advancements, barriers to clinical gene therapies remain. My research aims to address some of these barriers. The first topic of this thesis is the identification of gene editing loci (GELs), genomic regions where genetic information can be safely incorporated. Targeting these sites will improve gene editing safety and efficacy. Here, I demonstrate the functionality of targeting a proto-GEL on human chromosome 15 and discuss a script to identify novel GELs. The second topic focuses on the efficiency of gene insertion by DNA repair. I show that translesion DNA polymerase eta is important in the homology-directed repair pathway, which could help improve gene editing outcomes. I also reveal a potentially novel role of this enzyme in another aspect of genome maintenance, highlighting the dual nature of HDR-affecting enzymes.