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.