Use of genomic NW_006880285.1 in CHO cell for stably expressing a protein
AssigneeJiangnan University
InventorsJian Jin, Yun Chen, Huazhong Li, Songtao Zhou, Zuoying Duan, Xiaohai Gong
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Abstract
Use of genomic NW_006880285.1 in CHO cell for stably expressing a protein is disclosed. The certain site in CHO cell genome for stably expressing a protein is positioned at a base of No. 1235357 in a CHO cell gene NW_006880285.1; a sequence of 5′ NNNNNNNNNNNNNNNNNNNNNGG3′ that can be identified by CRISPR/Cas9 technology and positioned in a base range of No. 1235284-1235429 around the certain site is a target sequence. Various of protein genes are introduced into a fixed site in CHO cell genome, and expressed stably in the present disclosure.
Claims13
A method for stably expressing a protein at a predetermined site in a Chinese hamster ovary (CHO) cell genome, wherein the predetermined site in the CHO cell genome for stably expressing the protein is positioned at a base of No. 1235357 in a CHO cell gene NW_006880285.1;
- the method comprises:
- identifying a nucleotide sequence of SEQ ID NO:
- 15 in a base range of No. 1235284-1235429 in the CHO cell gene NW_006880285.1, the base range having the nucleotide sequence of SEQ ID NO:
- 12, around the predetermined site by CRISPR/Cas9 technology as a target sequence,
- transforming the CHO cell with a recombinant donor carrier containing the target sequence to obtain a recombinant CHO cell;
- culturing the recombinant CHO cell and collecting the supernatant to detect the expression level, and adapting an adherent recombinant CHO cell to a suspension culture;
- and
- culturing the adapted recombinant CHO cell.
Dependencies are inferred from the claim text.