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Last Updated: March 29, 2024

Claims for Patent: 9,567,578


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Summary for Patent: 9,567,578
Title:Cell line containing a knockout of the glutamine synthetase (GS) gene and a method of producing target proteins using a GS knockout HEK293 cell line
Abstract: The present invention relates to a novel GS (glutamine synthetase) gene knock out transgenic HEK293 (human embryonic kidney 293) cell line and a production method of a target protein using the said transgenic HEK293 cell line. Particularly, the present inventors eliminated the expression of GS in the HEK293 cells in order to overcome a barrier of the cell line selection caused by the over-expression of GS, for producing a target protein by GS/MSX system, by which the efficiency of the cell line selection for the high production of a target protein would be increased and accordingly the protein production by the selected cell line would be increased, suggesting that the human originated transgenic HEK293 cell line could be efficiently used for the production of a target protein.
Inventor(s): Lee; Gyun-Min (Daejeon, KR), Yu; Da Young (Daejeon, KR), Noh; Soo Min (Daejeon, KR)
Assignee: Korea Advanced Institute of Science and Technology (Daejeon, KR)
Application Number:15/138,165
Patent Claims:1. A glutamine synthetase (GS) knockout transgenic HEK293 cell line comprising one or more mutations of the nucleotide sequence of SEQ ID NO:1 within the 4.sup.th exon of the gs gene.

2. The GS knockout transgenic HEK293 cell line according to claim 1, wherein the cell line is the transgenic HEK293 cell line on deposit at the Korean Collection for Type Cultures (KCTC) under accession number KCTC12978BP.

3. The GS knockout transgenic HEK293 cell line according to claim 1, wherein the one or more mutations is an addition of an adenine nucleotide to the nucleotide sequence of SEQ ID NO:1 or a deletion of 11 nucleotides from the nucleotide sequence of SEQ ID NO:1.

4. The GS knockout transgenic HEK293 cell line according to claim 1, wherein the cell line is originated from HEK293 cells.

5. The GS knockout transgenic HEK293 cell line according to claim 4, wherein the HEK293 cell is selected from the group consisting of HEK293E, HEK293.sus, and HEK293T.

6. A method for producing a target protein comprising the following steps: 1) constructing a transformed HEK293 cell line manipulated to express a target protein by introducing a vector containing a gene encoding the target protein into the GS (glutamine synthetase) knockout transgenic HEK293 cell line of claim 1; 2) culturing the cell line prepared in step 1); and 3) separating and purifying the target protein produced in the cell line of step 2).

7. The method for producing a target protein according to claim 6, wherein the GS knockout transgenic HEK293 cell line of step 1) is the transgenic HEK293 cell line on deposit at the Korean Collection for Type Cultures (KCTC) under accession number KCTC12978BP.

8. The method for producing a target protein according to claim 6, wherein the target protein is selected from the group consisting of soluble TNF receptor, soluble IL-4 receptor, soluble IL-1 type II receptor, soluble CD40 ligand, CD39, CD30, CD27, TEK/ORK, IL-15 receptor, GM-CSF, RANKL, RANK, TRAIL, soluble TRAIL receptor, tissue plasminogen activator, factor VIII, factor IX, apolipoprotein E, apolipoprotein A-I, IL-2 receptor, IL-2 antagonist, alpha-1 antitrypsin, growth hormone, insulin-like growth factors, parathyroid hormone, interferon, monoclonal antibody (mAb), erythropoietin (EPO), thrombopoietin (TPO), and Fc containing fusion protein.

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