CPC A61K 35/17 (2013.01) [C07K 14/4718 (2013.01); C07K 14/70503 (2013.01); C07K 14/7051 (2013.01); C07K 14/7158 (2013.01); C12N 5/0636 (2013.01); C12N 9/22 (2013.01); C12N 9/96 (2013.01); C12N 15/113 (2013.01); C12N 15/907 (2013.01); C12N 15/87 (2013.01); C12N 2310/20 (2017.05); C12N 2510/00 (2013.01)] | 25 Claims |
1. A method of genomically modifying a population of human immune cells, the method comprising contacting ex vivo or in vitro the population of human immune cells with:
a) a polynucleic acid that encodes an exogenous T cell receptor or a chimeric antigen receptor, wherein said exogenous T cell receptor or said chimeric antigen receptor binds a cancer antigen;
b) a ribonuclear protein complex, wherein the ribonuclear protein complex comprises:
i) a guide RNA that binds a first gene locus within said human immune cells; and
ii) a Cas protein; and
c) a polynucleic acid that encodes a homologous recombination enhancer,
wherein the population of human immune cells are synchronized prior to said contacting with the ribonuclear protein complex, wherein the ribonuclear protein complex performs a genomic disruption within a target sequence of the first gene locus that comprises any one of SEQ ID NOS: 75-86, thereby generating genomically modified human immune cells, wherein the genomic disruption reduces or eliminates production of a protein encoded by the first gene locus as compared to a comparable human immune cell lacking the genomic disruption in the first gene locus, wherein the homologous recombination enhancer suppresses non-homologous end joining, and wherein said genomically modified human immune cells express said exogenous T cell receptor or chimeric antigen receptor.
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