Disruption of the gene C12orf35 leads to increased productivities in recombinant CHO cell lines
| Autor: | Genevieve Kuntzelmann, Bernd Voedisch, Panagiotis Mantas, Barbara Brannetti, Tatjana Rauschert, Nadine Lageyre, Sabine Geisse, Daniel Piehlmaier, Anett Ritter, Mevion Oertli, Holger Laux, Thomas Jostock |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Messenger RNA biology Chinese hamster ovary cell Bioengineering Immunoglobulin light chain Applied Microbiology and Biotechnology Phenotype Molecular biology law.invention 03 medical and health sciences 030104 developmental biology law Cell culture Dihydrofolate reductase biology.protein Recombinant DNA Gene Biotechnology |
| Zdroj: | Biotechnology and Bioengineering. 113:2433-2442 |
| ISSN: | 0006-3592 |
| DOI: | 10.1002/bit.26009 |
| Popis: | Recently, we reported that the loss of a telomeric region of chromosome 8 in Chinese Hamster Ovary (CHO) cells correlates with higher recombinant productivities. New cell lines lacking this region, called CHO-C8DEL, showed several advantages during cell line generation and for the production of recombinant proteins (Ritter et al., 2016, Biotechnol Bioeng). Here, we performed knock-down and knock-out experiments of genes located within this telomeric region of chromosome 8 to identify the genes causing the observed phenotypes of CHO-C8DEL cell lines. We present evidence that loss or reduced expression of the gene C12orf35 is responsible for higher productivities and shorter recovery times during selection pressure. These effects are mediated by increased levels of mRNA of the exogenes heavy chain (HC) and light chain (LC) as well as dihydrofolate reductase (DHFR) and neomycin phosphotransferase (Neo) during the stable expression of antibodies. Biotechnol. Bioeng. 2016;113: 2433-2442. © 2016 Wiley Periodicals, Inc. |
| Databáze: | OpenAIRE |
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