Stable overexpression of miR-17 enhances recombinant protein production of CHO cells

被引:61
作者
Jadhav, Vaibhav [1 ]
Hackl, Matthias [1 ]
Klanert, Gerald [1 ,2 ]
Bort, Juan A. Hernandez [1 ,2 ]
Kunert, Renate [1 ]
Grillari, Johannes [1 ]
Borth, Nicole [1 ,2 ]
机构
[1] Univ Nat Resources & Life Sci, Dept Biotechnol, Vienna, Austria
[2] Austrian Ctr Ind Biotechnol GmbH, Graz, Austria
关键词
Chinese hamster ovary (CHO) cells; microRNA (miRNA); miR-17; miR-92a; miR17-92a cluster; Productivity; MAMMALIAN MICRORNAS; MESSENGER-RNA; EXPRESSION; MIRNA; PATHWAY; GROWTH; GENE; TEMPERATURE; INHIBITION; BIOGENESIS;
D O I
10.1016/j.jbiotec.2014.01.032
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
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. (C) 2014 The Authors. Published by Elsevier B.V. All rights reserved.
引用
收藏
页码:38 / 44
页数:7
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