	US 11,859,207 B2
	Artificial antigen-presenting cell prepared from HLA-null cell line by using multiplex CRISPR-CAS9 system and use thereof
Tai Gyu Kim, Seoul (KR); Hyun Jung Sohn, Seoul (KR); and Cheol Hwa Hong, Seoul (KR)
Assigned to The Catholic Uiversity of Korea Industry-Academic Cooperation Foundation, Seoul (KR)
Appl. No. 16/465,933
Filed by THE CATHOLIC UNIVERSITY OF KOREA INDUSTRY-ACADEMIC COOPERATION FOUNDATION, Seoul (KR)
PCT Filed Dec. 1, 2017, PCT No. PCT/KR2017/014027 § 371(c)(1), (2) Date May 31, 2019, PCT Pub. No. WO2018/101796, PCT Pub. Date Jun. 7, 2018.
Claims priority of application No. 10-2016-0163509 (KR), filed on Dec. 2, 2016.
Prior Publication US 2019/0309260 A1, Oct. 10, 2019
Int. Cl. C12N 5/078 (2010.01); A61K 35/17 (2015.01); C07K 14/74 (2006.01); C12N 5/0783 (2010.01); C12N 9/22 (2006.01); C12N 15/11 (2006.01); C12N 15/85 (2006.01); C12N 15/63 (2006.01)

CPC C12N 5/0634 (2013.01) [A61K 35/17 (2013.01); C07K 14/70539 (2013.01); C12N 5/0638 (2013.01); C12N 9/22 (2013.01); C12N 15/11 (2013.01); C12N 15/63 (2013.01); C12N 15/85 (2013.01); C12N 2310/20 (2017.05); C12N 2502/99 (2013.01); C12N 2800/80 (2013.01)]

1 Claim

1. A method to make one or more antigen-presenting cells, comprising:

a) eliminating HLA-A, HLA-B and HLA-C in one or more 293T cells using a CRISPR-Cas9 system to make one or more HLA-null 293T cells,

wherein the CRISPR-Cas9 system comprises single guide RNAs(sgRNAs) targeting each of:

HLA-A exon 2(SEQ ID NO: 1) and exon 3(SEQ ID NO: 2),

HLA-B exon 2(SEQ ID NO: 3) and exon 3(SEQ ID NO: 4),

HLA-C exon 2(SEQ ID NO: 5) and exon 3(SEQ ID NO: 6); and

b) introducing a co-stimulatory molecule group into the one or more HLA-null 293T cells to make one or more antigen-presenting cells,

wherein the co-stimulatory molecule is selected from the group consisting of: CD80; CD83; CD54; CD32; 4-1BBL; and CD70.