Technology field: Genome editing, CRISPR-Cas systems, molecular biology
A method for editing the genome of a eukaryotic cell using a CRISPR-Cas9 system comprising: (1) a guide RNA (gRNA) engineered to include a spacer sequence complementary to a target DNA sequence within the eukaryotic cell's genome; (2) a Cas9 endonuclease capable of forming a complex with the gRNA and introducing a double-strand break at the target DNA sequence; and (3) optionally, a donor DNA template for homology-directed repair. The method enables precise, programmable editing of genomic loci in human and other mammalian cells by directing the Cas9-gRNA ribonucleoprotein complex to specified chromosomal positions via Watson-Crick base pairing between the spacer and the protospacer adjacent to a PAM sequence. Applications include correction of genetic mutations, gene knockouts, and insertion of exogenous sequences.