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

Jadhav, Vaibhav; Hackl, Matthias; Klanert, Gerald; Bort, Juan A. Hernández; Kunert, Renate; Grillari, Johannes; Borth, Nicole

doi:10.1016/j.jbiotec.2014.01.032

Cited by 69 publications

(66 citation statements)

References 46 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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“…Jadhav V et al reported a similar result and designed an experiment to explain this phenomenon [34]. Their results demonstrated that transient positive effects on growth could also be observed during early exponential culture.…”

Section: Discussionmentioning

confidence: 76%

Negative feedback between TAp63 and Mir-133b mediates colorectal cancer suppression

Dai

Zhang

et al. 2016

Oncotarget

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BackgroundTAp63 is known as the most potent transcription activator and tumor suppressor. microRNAs (miRNAs) are increasingly recognized as essential components of the p63 pathway, mediating downstream post-transcriptional gene repression. The aim of present study was to investigate a negative feedback loop between TAp63 and miR-133b.ResultsOverexpression of TAp63 inhibited HCT-116 cell proliferation, apoptosis and invasion via miR-133b. Accordingly, miR-133b inhibited TAp63 expression through RhoA and its downstream pathways. Moreover, we demonstrated that TAp63/miR-133b could inhibit colorectal cancer proliferation and metastasis in vivo and vitro.Materials and MethodsWe evaluated the correlation between TAp63 and miR-133b in HCT-116 cells and investigated the roles of the TAp63/miR-133b feedback loop in cell proliferation, apoptosis and metastasis via MTT, flow cytometry, Transwell, and nude mouse xenograft experiments. The expression of TAp63, miR-133b, RhoA, α-tubulin and Akt was assessed via qRT-PCR, western blot and immunofluorescence analyses. miR-133b target genes were identified through luciferase reporter assays.ConclusionsmiR-133b plays an important role in the anti-tumor effects of TAp63 in colorectal cancer. miR-133b may represent a tiemolecule between TAp63 and RhoA, forming a TAp63/miR-133b/RhoA negative feedback loop, which could significantly inhibit proliferation, apoptosis and metastasis.

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“…From these five, miR-2110 is a novel miRNA that has been identified but not studied (Zhu et al 2009). The other four miRNAs (miR-429, miR-18a-5p, miR-22-5p and miR-22-3p) have been associated with cancer research in which they have exhibited contradicting effects on cell proliferation, cell growth, and protein production depending on the cell type and stage of cell development (Jadhav et al 2014; Lang et al 2014; Liu et al 2014; Song et al 2013; Song et al 2014; Takakura et al 2008; Xiong et al 2010). For example, miR-429, a member of the miR-200 family, was shown to suppress tumor growth in human osteosarcoma(Liu et al 2014), while in non-small cell lung cancer (NSCLC), the same miRNA is suggested as a potential target for NSCLC due to its promotion of cell proliferation(Lang et al 2014).…”

Section: Discussionmentioning

confidence: 99%

“…This miR-17-92 cluster was studied in depth regarding its effect on recombinant EpoFc protein production in CHO cells. Although over-expression of the entire cluster decreased productivity while having no effect on cell growth, the over-expression of miR-17 and miR-92 were shown to increase production(Jadhav et al 2014). …”

Section: Discussionmentioning

confidence: 99%

“…MiRNAs have emerged as powerful tools for engineering cells with desirable properties (Jadhav et al 2013), such as improved protein production capabilities (Fischer et al 2014; Jadhav et al 2014; Loh et al 2014) and enhanced anti-apoptotic properties under stress conditions(Druz et al 2013a; Druz et al 2013b). MiRNAs are a novel class of small, non-coding RNAs that can simultaneously silence multiple genes by binding to their 3′-untranslated regions(3′-UTR)(Filipowicz et al 2008).…”

Section: Introductionmentioning

confidence: 99%

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MiRNA mimic screen for improved expression of functional neurotensin receptor from HEK 293 cells

Xiao

Chen

Betenbaugh

et al. 2015

Biotech & Bioengineering

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Obtaining adequate quantities of functional mammalian membrane proteins has been a bottleneck in their structural and functional studies because the expression of these proteins from mammalian cells is relatively low. To explore the possibility of enhancing expression of these proteins using miRNA, a stable T-REx-293 cell line expressing the neurotensin receptor type 1 (NTSR1), a hard-to-express G protein-coupled receptor (GPCR), was constructed. The cell line was then subjected to human miRNA mimic library screening. In parallel, an HEK293 cell line expressing luciferase was also screened with the same human miRNA mimic library. Five microRNA mimics: hsa-miR-22-5p, hsa-miR-18a-5p, hsa-miR-22-3p, hsa-miR-429 and hsa-miR-2110 were identified from both screens. They led to 48% increase in the expression of functional NTSR1 and to 239% increase of luciferase expression. These miRNAs were also effective in enhancing the expression of secreted glypican-3 hFc-fusion protein from HEK293 cells. The results indicate that these molecules may have a wide role in enhancing the production of proteins with biomedical interest.

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

Abstract: HighlightsTransient overexpression of miR-17 and miR-17–92 cluster enhanced growth rate.Biological effects of long term and stable overexpression of miRNAs in batch cultures were studied.Stable miR-17 engineered CHO cells had both improved growth rate and productivity.

Cited by 69 publications

References 46 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

Negative feedback between TAp63 and Mir-133b mediates colorectal cancer suppression

MiRNA mimic screen for improved expression of functional neurotensin receptor from HEK 293 cells

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