title: Stable overexpression of miR-17 enhances recombinant protein production of CHO cells.
abstract: miRNAs negatively regulate gene expression at post-transcriptional level, and consequently play an important role in the control of many cellular pathways. The use of miRNAs to engineer Chinese hamster ovary (CHO) cells is an emerging strategy to improve recombinant protein production. Here, we describe the effect of transient and stable miRNA overexpression on CHO cell phenotype. Using an established transient miRNA screening protocol, the effects of miR-17, miR-92a and cluster miR17-92a on CHO growth and protein productivity were studied and followed by analysis of cell pools with stable overexpression of these miRNAs. CHO cells stably engineered with miR-17 exhibited both enhanced growth performance and a 2-fold increase in specific productivity, which resulted in a 3-fold overall increase in EpoFc titer. While further studies of miRNA-mRNA interactions will be necessary to understand the molecular basis of this effect, these data provide valuable evidence for miR-17 as a cell engineering target to enhance CHO cell productivity.
authors: Department of Biotechnology, University of Natural Resources and Life Sciences, Vienna, Austria.
authors: Vaibhav
authors: V
authors: Jadhav
authors: Department of Biotechnology, University of Natural Resources and Life Sciences, Vienna, Austria.
authors: Matthias
authors: M
authors: Hackl
authors: Department of Biotechnology, University of Natural Resources and Life Sciences, Vienna, Austria; Austrian Centre of Industrial Biotechnology GmbH, Graz, Austria.
authors: Gerald
authors: G
authors: Klanert
authors: Department of Biotechnology, University of Natural Resources and Life Sciences, Vienna, Austria; Austrian Centre of Industrial Biotechnology GmbH, Graz, Austria.
authors: Juan A
authors: JA
authors: Hernandez Bort
authors: Department of Biotechnology, University of Natural Resources and Life Sciences, Vienna, Austria.
authors: Renate
authors: R
authors: Kunert
authors: Department of Biotechnology, University of Natural Resources and Life Sciences, Vienna, Austria.
authors: Johannes
authors: J
authors: Grillari
authors: Department of Biotechnology, University of Natural Resources and Life Sciences, Vienna, Austria; Austrian Centre of Industrial Biotechnology GmbH, Graz, Austria. Electronic address: nicole.borth@boku.ac.at.
authors: Nicole
authors: N
authors: Borth
mesh_terms: D000818:Animals; D016466:CHO Cells / Q000378:metabolism*; D060846:Cell Engineering / Q000379:methods; D049109:Cell Proliferation; D006224:Cricetinae; D003412:Cricetulus; D006801:Humans; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism*; D010641:Phenotype; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
chemical_list: D035683:MicroRNAs; D011994:Recombinant Proteins
keywords: Chinese hamster ovary (CHO) cells; Productivity; miR-17; miR-92a;; miR17-92a cluster; microRNA (miRNA)