Genome Editing: From Fundamental DNA Repair Mechanisms to Cutting-Edge Applications

Yinan Kan^1,2,3, Eric Hendrickson^1,3, George Church^1,2, Luhan Yang^1,2

¹ eGenesis Inc., 300 Technology Square, Cambridge, MA 02139, USA

² Department of Genetics, Harvard Medical School, Boston, MA 02115, USA

³ Biochemistry, Molecular Biology and Biophysics, University of Minnesota, Minneapolis, MN 55455, USA

Genome editing is the intentional alteration of genetic information in living cells or organisms. Since CRISPR/Cas9 was repurposed for genome editing, the “CRISPR Craze” is quickly bridging the genotype and phenotype worlds and transforming biological and biomedical research. Interestingly, CRISRP/Cas9 does not perform genome editing by itself, but it induces a site-specific genomic lesion and relies on the endogenous DNA repair pathways to introduce desired modifications to the genome.

In this presentation, I will walk through the DNA repair mechanisms of genome editing with a focus on the non-homologous end joining (NHEJ) and homology-directed repair (HDR) pathways. Then I will highlight some of advanced applications of the NHEJ and HDR pathways including multiplex gene knockout and marker-free long-tract gene conversion. Finally, I will introduce eGenesis’ recent efforts in generating the most extensively genome-edited large animals with the aim of providing safe and effective porcine organs for human transplantation.