OPTIMIZING GENE TARGETING METHODOLOGIES USING CRISPR/CAS RIBONUCLEOPROTEINS IN PORCINE SOMATIC CELLS

Abstract

The pig is one of the most valuable animal models for agricultural and biomedical research. In this thesis, we sought to standardize procedures for routine gene targeting and demonstrate that it is possible to create site-specific knockins of short DNA sequences in a locus of our interest, specifically downstream of a pig insulin gene and a ubiquitously expressed COL1A gene in pigs. Our study successfully established and optimized protocol for CRISPR/Cas mediated knockin in porcine somatic cells, while simultaneously creating a stable platform for future transgene integration and stepping forward to perform more efficient porcine transgenesis in the future.