title: Overexpression of microRNAs enhances recombinant protein production in Chinese hamster ovary cells.
abstract: MicroRNAs (miRNAs) are short, non-coding RNAs that can negatively regulate expression of multiple genes at post-transcriptional levels. Using miRNAs to target multiple genes and pathways is a promising cell-engineering strategy to increase recombinant protein production in mammalian cells. Here, we identified miRs-17, -19b, -20a, and -92a to be differentially expressed between high- and low- monoclonal antibody-producing Chinese hamster ovary (CHO) cell clones using next-generation sequencing and quantitative real-time PCR. These miRNAs were stably overexpressed individually and in combination in a high-producing clone to assess their effects on CHO cell growth, recombinant protein productivity and product quality. Stably transfected pools demonstrated 24-34% increases in specific productivity (qP) and 21-31% increases in titer relative to the parental clone, without significant alterations in proliferation rates. The highest protein-producing clones isolated from these pools exhibited 130-140% increases in qP and titer compared to the parental clone, without major changes in product aggregation and N-glycosylation profile. From our clonal data, correlations between enhanced qP/titer and increased levels of miRs-17, -19b, and -92a were observed. Our results demonstrate the potential of miRs-17, -19b, and -92a as cell-engineering targets to increase recombinant protein production in mammalian cells.
authors.affiliation: Bioprocessing Technology Institute, Agency for Science, Technology and Research (A*STAR), Singapore. loh_wan_ping@bti.a-star.edu.sg.
authors.forename: Wan Ping
authors.initials: WP
authors.lastname: Loh
authors.forename: Bernard
authors.initials: B
authors.lastname: Loo
authors.forename: Lihan
authors.initials: L
authors.lastname: Zhou
authors.forename: Peiqing
authors.initials: P
authors.lastname: Zhang
authors.forename: Dong-Yup
authors.initials: DY
authors.lastname: Lee
authors.forename: Yuansheng
authors.initials: Y
authors.lastname: Yang
authors.forename: Kong Peng
authors.initials: KP
authors.lastname: Lam
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics; D016466:CHO Cells; D060846:Cell Engineering / Q000379:methods; D002460:Cell Line; D049109:Cell Proliferation / Q000235:genetics; D003412:Cricetulus; D035683:MicroRNAs / Q000235:genetics*; D011994:Recombinant Proteins / Q000235:genetics*
chemical_list: D000911:Antibodies, Monoclonal; D035683:MicroRNAs; D011994:Recombinant Proteins
keywords: CHO cells; Cell engineering; Recombinant protein production; miR-17-92 cluster; microRNA profiling