title: Disruption of the gene C12orf35 leads to increased productivities in recombinant CHO cell lines.
abstract: 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.
authors.affiliation: Novartis Institutes for BioMedical Research, Basel, Switzerland. anett.ritter@novartis.com.|Novartis Pharma AG, Integrated Biologics Profiling Unit, Werk Klybeck Postfach CH-4002, Basel, Switzerland. anett.ritter@novartis.com.
authors.forename: Anett
authors.initials: A
authors.lastname: Ritter
authors.affiliation: Novartis Pharma AG, Cell and Gene Therapies, Basel, Switzerland.
authors.forename: Tatjana
authors.initials: T
authors.lastname: Rauschert
authors.affiliation: Novartis Institutes for BioMedical Research, Basel, Switzerland.|Novartis Pharma AG, Integrated Biologics Profiling Unit, Werk Klybeck Postfach CH-4002, Basel, Switzerland.
authors.forename: Mevion
authors.initials: M
authors.lastname: Oertli
authors.affiliation: Novartis Pharma AG, Integrated Biologics Profiling Unit, Werk Klybeck Postfach CH-4002, Basel, Switzerland.
authors.forename: Daniel
authors.initials: D
authors.lastname: Piehlmaier
authors.affiliation: Novartis Pharma AG, Integrated Biologics Profiling Unit, Werk Klybeck Postfach CH-4002, Basel, Switzerland.
authors.forename: Panagiotis
authors.initials: P
authors.lastname: Mantas
authors.affiliation: Novartis Institutes for BioMedical Research, Basel, Switzerland.
authors.forename: Genevieve
authors.initials: G
authors.lastname: Kuntzelmann
authors.affiliation: Novartis Institutes for BioMedical Research, Basel, Switzerland.
authors.forename: Nadine
authors.initials: N
authors.lastname: Lageyre
authors.affiliation: Novartis Institutes for BioMedical Research, Basel, Switzerland.
authors.forename: Barbara
authors.initials: B
authors.lastname: Brannetti
authors.affiliation: Novartis Institutes for BioMedical Research, Basel, Switzerland.
authors.forename: Bernd
authors.initials: B
authors.lastname: Voedisch
authors.affiliation: Novartis Institutes for BioMedical Research, Basel, Switzerland.
authors.forename: Sabine
authors.initials: S
authors.lastname: Geisse
authors.affiliation: Novartis Pharma AG, Integrated Biologics Profiling Unit, Werk Klybeck Postfach CH-4002, Basel, Switzerland.
authors.forename: Thomas
authors.initials: T
authors.lastname: Jostock
authors.affiliation: Novartis Pharma AG, Integrated Biologics Profiling Unit, Werk Klybeck Postfach CH-4002, Basel, Switzerland. holger.laux@novartis.com.
authors.forename: Holger
authors.initials: H
authors.lastname: Laux
mesh_terms: D000818:Animals; D016466:CHO Cells / Q000166:cytology / Q000502:physiology*; D003412:Cricetulus; D055785:Gene Knockdown Techniques; D024861:Genetic Enhancement / Q000379:methods*; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D015854:Up-Regulation / Q000235:genetics
chemical_list: D011994:Recombinant Proteins
keywords: Chinese hamster ovary cells; cell line engineering; chromosomal deletion; mRNA; productivity; recombinant gene expression