US 11,925,664 B2
Intracellular genomic transplant and methods of therapy
Branden Moriarity, Shoreview, MN (US); Beau Webber, Coon Rapids, MN (US); David Largaespada, Mounds View, MN (US); Modassir Choudhry, New York, NY (US); and Steven A. Rosenberg, Potomac, MD (US)
Assigned to INTIMA BIOSCIENCE, INC., New York, NY (US); REGENTS OF THE UNIVERSITY OF MINNESOTA, Minneapolis, MN (US); and THE UNITED STATES OF AMERICA, AS REPRESENTED BY THE SECRETARY, DEPARTMENT OF HEALTH AND HUMAN SERVICES, Bethesda, MD (US)
Filed by Regents of the University of Minnesota, Minneapolis, MN (US); Intima Bioscience, Inc., New York, NY (US); and The U.S.A., as Represented by the Secretary, Department of Health and Human Services, Bethesda, MD (US)
Filed on Jun. 12, 2020, as Appl. No. 16/900,368.
Application 16/900,368 is a continuation of application No. 16/180,867, filed on Nov. 5, 2018.
Application 16/180,867 is a continuation of application No. 15/224,151, filed on Jul. 29, 2016, granted, now 10,166,255, issued on Jan. 1, 2019.
Claims priority of provisional application 62/360,245, filed on Jul. 8, 2016.
Claims priority of provisional application 62/330,464, filed on May 2, 2016.
Claims priority of provisional application 62/295,670, filed on Feb. 16, 2016.
Claims priority of provisional application 62/286,206, filed on Jan. 22, 2016.
Claims priority of provisional application 62/232,983, filed on Sep. 25, 2015.
Claims priority of provisional application 62/199,905, filed on Jul. 31, 2015.
Prior Publication US 2020/0353001 A1, Nov. 12, 2020
Int. Cl. A61K 48/00 (2006.01); A61K 35/17 (2015.01); C07K 14/47 (2006.01); C07K 14/705 (2006.01); C07K 14/715 (2006.01); C07K 14/725 (2006.01); C12N 5/0783 (2010.01); C12N 9/22 (2006.01); C12N 9/96 (2006.01); C12N 15/07 (2006.01); C12N 15/113 (2010.01); C12N 15/90 (2006.01); C12N 15/87 (2006.01)
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.