UC San Diego

With the advent of recombinant DNA technology in the 1970s, the idea of using genetherapies to treat human genetic diseases captured the interest and imagination of scientists around the world. Years later, enabled largely by the development of CRISPR-based genome editing tools, the field has exploded, with academic labs, startup biotechnology companies, and large pharmaceutical corporations working in concert to develop life-changing therapeutics. Prime editing is a new genome editing methodology that utilizes novel intermediates. How the cell processes these intermediates into desired genome editing outcomes has yet to be fully characterized. In Chapter 2 and 3, we detail our findings which aim to provide insights into the mechanisms governing prime editing and reveal this new technology to be more ubiquitous than traditional genome editing tools that rely on S- and G2-phase dependent DNA repair pathways. Base editing, first reported in 2016, is capable of installing C•G to T•A and A•T to G•C point mutations, while largely circumventing some of the pitfalls of traditional CRISPR/Cas9 gene editing. Despite their youth, these technologies have been widely used by both academic labs and therapeutics-based companies. In Chapter 4, we detail work aimed at addressing two of the main pitfalls of base editors: limited point mutation installation capability and incompatibility with optimal viral therapeutic delivery vehicles. Finally, in Chapter 5, non-scientific development garnered throughout the primary author and researcher’s tenure is briefly discussed as this work has also led to both personal and professional advances.