	US 12,188,036 B2
	Expression cassette for production of high-expression and high-functionality target protein and use thereof
Man Su Kim, Incheon (KR); Min Soo Kim, Incheon (KR); Jong Moon Cho, Incheon (KR); and Shin Jae Chang, Incheon (KR)
Assigned to CELLTRION INC., Incheon (KR)
Appl. No. 16/755,453
Filed by CELLTRION INC., Incheon (KR)
PCT Filed Oct. 11, 2018, PCT No. PCT/KR2018/011953 § 371(c)(1), (2) Date Apr. 10, 2020, PCT Pub. No. WO2019/074292, PCT Pub. Date Apr. 18, 2019.
Claims priority of application No. 10-2017-0129969 (KR), filed on Oct. 11, 2017.
Prior Publication US 2020/0299721 A1, Sep. 24, 2020
Int. Cl. C12N 15/85 (2006.01); C07K 16/00 (2006.01); C12N 15/11 (2006.01); C12N 15/113 (2010.01)

CPC C12N 15/85 (2013.01) [C12N 15/111 (2013.01); C12N 2830/42 (2013.01); C12N 2840/44 (2013.01)]

8 Claims

1. An expression cassette for production of a target protein, comprising a single promoter, a polynucleotide coding for the target protein, a poly A sequence, and a polynucleotide sequence encoding an intronic RNA sequence,

wherein the intronic RNA sequence comprises a splicing donor, a branch, a splicing acceptor, and a target gene expression regulation RNA sequence;

wherein the target protein is an antibody or a fragment thereof;

wherein the single promoter transcriptionally links the polynucleotide coding for the target protein and the polynucleotide sequence encoding the intronic RNA;

and wherein the target gene expression regulation RNA sequence comprises one or more shRNA (small hairpin RNA) sequences selected from the group consisting of intronic FUT8 (Alpha-1,6-fucosyltransferase) shRNA represented by SEQ ID NO: 24, intronic HDAC5 (histone deacetylase 5) shRNA represented by any one of SEQ ID NOS: 25 to 27, intronic LDHA (lactate dehydrogenase A) shRNA represented by any one of SEQ ID NOS: 28 to 30, and intronic DHFR (dihydrofolate reductase) shRNA represented by any one of SEQ ID NOS: 31 to 33.