Engineering CHO cell growth and recombinant protein productivity by overexpression of miR-7

Barron, Niall; Kumar, Niraj; Sánchez, Nilda; Doolan, Padraig; Clarke, Colin; Meleady, Paula; O'Sullivan, Fin; Clynes, Martin

doi:10.1016/j.jbiotec.2010.12.005

Cited by 106 publications

(72 citation statements)

References 36 publications

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“…Subsequent studies were focused on providing insights into the importance of miRNAs for molecular pathways relevant to cell culture engineering, such as growth (Jadhav et al, 2014), apoptosis (Druz et al, 2011), and protein secretion (Barron et al, 2011; Loh et al, 2014). …”

Section: Introductionmentioning

confidence: 99%

Annotation of additional evolutionary conserved microRNAs in CHO cells from updated genomic data

Diendorfer

Hackl

Klanert

et al. 2015

Biotech & Bioengineering

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MicroRNAs are small non‐coding RNAs that play a critical role in post‐transcriptional control of gene expression. Recent publications of genomic sequencing data from the Chinese Hamster (CGR) and Chinese hamster ovary (CHO) cells provide new tools for the discovery of novel miRNAs in this important production system. Version 20 of the miRNA registry miRBase contains 307 mature miRNAs and 200 precursor sequences for CGR/CHO. We searched for evolutionary conserved miRNAs from miRBase v20 in recently published genomic data, derived from Chinese hamster and CHO cells, to further extend the list of known miRNAs. With our approach we could identify several hundred miRNA sequences in the genome. For several of these, the expression in CHO cells could be verified from multiple next‐generation sequencing experiments. In addition, several hundred unexpressed miRNAs are awaiting further confirmation by testing for their transcription in different Chinese hamster tissues. Biotechnol. Bioeng. 2015;112: 1488–1493. © 2015 The Authors. Biotechnology and Bioengineering Published by Wiley Periodicals, Inc.

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Section: Introductionmentioning

confidence: 99%

Annotation of additional evolutionary conserved microRNAs in CHO cells from updated genomic data

Diendorfer

Hackl

Klanert

et al. 2015

Biotech & Bioengineering

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“…Target-miR interactions have been demonstrated in multiple cell types and disease settings, particularly in mouse and human, but miRNA targets in CHO cells are still being elucidated (Meleady et al, 2011). Our results show for the first time that inhibition of miR let-7a in CHO cells led to changes in multiple genes/proteins previously associated with cell cycle control/proliferation and apoptosis, stress response, cellular metabolism, and protein transcription/translation.…”

Section: Discussionmentioning

confidence: 66%

“…Accordingly, investigators have begun to examine the role of microRNAs in CHO cell cultures (Muller et al, 2008;Gammell et al, 2007;Hackl et al, 2010; primarily through analysis of alterations in endogenous miRNAs that occur throughout production culture. A small number of studies have also explored the effect of ectopically expressed miRs or anti-miRs on CHO cells Meleady et al, 2011;Druz et al, 2011, Strotbek et al, 2013 however, broader exploration and careful characterization of the ability to use miRNA to engineer CHO cell lines is required before this technology can be implemented routinely to increase production of therapeutic biologics.…”

Section: Introductionmentioning

confidence: 99%

Inhibition of microRNA-let-7a Increases the Specific Productivity of Antibody-Producing CHO Cell Lines

Greenlees¹,

Georgantas²,

Zhu³

et al. 2014

gab

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Chinese hamster ovary cells (CHO) are the preferred cell line for the production of recombinant biopharmaceuticals, which constitutes a multi-billion dollar global market. Major challenges to improving protein productivity of CHO in large-scale production cultures include growth level, cellular stress, and translation rate. Because microRNA (miR, miRNA) can simultaneously perturb multiple pathways by inhibiting translation or destabilizing different mRNAs, we explored their utility to extend the cell growth phase or alter protein production per cell (specific productivity) with the goal of enhancing current optimization techniques to increase the production capacity of CHO cell cultures. To investigate the effect of altered microRNA expression on CHO cell viability and specific productivity, two clinically relevant antibody-producing CHO cell lines were stably transduced with lentiviral vectors encoding nine different miRNAs or anti-miRNAs based on their potential involvement in pathways critical for recombinant protein production. Inhibition of miR let-7a led to a 50%~68% increase in specific productivity in two recombinant antibody-producing cell lines. Furthermore, following miR let-7a inhibition, we identified increased expression of its targets HMGA2, MYC, NF2, NIRF, RAB40C, and eIF4a which are important mediators of apoptosis, protein translation, and cellular metabolism. Overall, this work provides proof of concept that exogenous microRNA modifications can positively affect specific productivity of CHO cell cultures and highlights the potential of miR let-7a to have a broad impact on the complex biological functions necessary for improving the capabilities of CHO cell lines.

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“…Whereas, inhibition of UBA52 didn't showed similar effects on cellular health as expression of only ~7% mRNAs was observed to be altered suggesting transcript-specific translation by the UBA52 [39]. This could be important for recombinant protein production as low temperature has been believed to cause transcript-specific translation and hence improve overall yield from the cultures [18,39,40]. USP5 is involved in disassembly of conjugated ubiquitin to maintain proteasome function and its availability for other functions [41].…”

Section: Functional Analysis Of Microvesicular Proteinsmentioning

confidence: 99%

Exploring Packaged Microvesicle Proteome Composition of Chinese Hamster Ovary Secretome

2016

J Bioprocess Biotech

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Engineering CHO cell growth and recombinant protein productivity by overexpression of miR-7

Cited by 106 publications

References 36 publications

Annotation of additional evolutionary conserved microRNAs in CHO cells from updated genomic data

Annotation of additional evolutionary conserved microRNAs in CHO cells from updated genomic data

Inhibition of microRNA-let-7a Increases the Specific Productivity of Antibody-Producing CHO Cell Lines

Exploring Packaged Microvesicle Proteome Composition of Chinese Hamster Ovary Secretome

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