	US 12,275,963 B2
	Artificially manipulated immune cell
Seok Joong Kim, Seoul (KR); Yoon-Young Kim, Seoul (KR); Ho-Sung Yu, Gyeonggi-do (KR); In-Young Jung, Gyeonggi-do (KR); and Jung Min Lee, Gyeongsangbuk-do (KR)
Assigned to TOOLGEN INCORPORATED, Seoul (KR)
Appl. No. 16/611,383
Filed by TOOLGEN INCORPORATED, Seoul (KR)
PCT Filed May 8, 2018, PCT No. PCT/KR2018/005284 § 371(c)(1), (2) Date Nov. 6, 2019, PCT Pub. No. WO2018/208067, PCT Pub. Date Nov. 15, 2018.
Claims priority of provisional application 62/595,159, filed on Dec. 6, 2017.
Claims priority of provisional application 62/502,822, filed on May 8, 2017.
Claims priority of application No. PCT/KR2017/008835 (WO), filed on Aug. 14, 2017.
Prior Publication US 2021/0147798 A1, May 20, 2021
Int. Cl. A61K 39/00 (2006.01); C07K 14/705 (2006.01); C07K 16/28 (2006.01); C12N 5/078 (2010.01); C12N 5/0783 (2010.01); C12N 9/22 (2006.01); C12N 15/10 (2006.01); C12N 15/113 (2010.01); C12N 15/115 (2010.01); C12N 15/117 (2010.01); C12N 15/87 (2006.01); C07K 14/725 (2006.01)

CPC C12N 9/22 (2013.01) [A61K 39/4611 (2023.05); A61K 39/4631 (2023.05); A61K 39/4644 (2023.05); C07K 14/705 (2013.01); C07K 16/2863 (2013.01); C12N 5/0634 (2013.01); C12N 5/0636 (2013.01); C12N 15/102 (2013.01); C12N 15/113 (2013.01); C12N 15/115 (2013.01); C12N 15/117 (2013.01); C12N 15/87 (2013.01); C12Y 207/01107 (2013.01); A61K 2239/31 (2023.05); A61K 2239/38 (2023.05); C07K 14/7051 (2013.01); C07K 2317/622 (2013.01); C07K 2319/03 (2013.01); C12N 2501/515 (2013.01)]

13 Claims

1. A manipulated human T cell with an enhanced immune activity comprising:

an engineered genome comprising an engineered endogenous DGKA gene and an engineered endogenous DGKZ gene; and

at least one artificial receptor and/or nucleic acid encoding the artificial receptor,

wherein the engineered endogenous DGKA gene comprises a first artificial modification,

wherein the first artificial modification is a first indel by CRISPR/Cas gene editing and induced in a region of exon 7 of the wild-type DGKA gene, wherein the engineered endogenous DGKA gene does not comprise any further modification other than the first artificial modification,

wherein the engineered endogenous DGKZ gene comprises a second artificial modification,

wherein the second artificial modification is a second indel by CRISPR/Cas gene editing and induced in a region of exon 3 of the wild-type DGKZ gene, wherein the engineered endogenous DGKZ gene does not comprise any further modification other than the second artificial modification,

wherein the first artificial modification causes at least one of a reduced expression of engineered endogenous DGKA gene and a functional impairment of the protein expressed from the engineered endogenous DGKA gene, and the second artificial modification causes at least one of a reduced expression of engineered endogenous DGKZ gene and a functional impairment of the protein expressed from the engineered endogenous DGKZ gene, in a manner that the manipulated human T cell has the enhanced immune activity.