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Cited by 73Viral Infectious Diseases and Gene Expression in InsectsMicroRNA in disease regulationCell death mechanisms and regulation

Stable inhibition of mmu-miR-466h-5p improves apoptosis resistance and protein production in CHO cells

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Druz, Aliaksandr, Son, Young–Jin, Betenbaugh, Michael, Shiloach, Joseph·2013·Metabolic Engineering
DOI 10.1016/j.ymben.2012.12.004PMC PMC3644357
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Abstract

MiRNAs have been shown to be involved in regulation of multiple cellular processes including apoptosis. Since a single miRNA can affect the expression of several genes, the utilization of miRNAs for apoptosis engineering in mammalian cells can be more efficient than the conventional approach of manipulating a single gene. Mmu-miR-466h-5p was previously shown to have a pro-apoptotic role in CHO cells by reducing the expression of several anti-apoptotic genes and its transient inhibition delayed both the activation of Caspase-3/7 and the loss of cell viability. The present study evaluates the effect of stable inhibition of mmu-miR-466h-5p in CHO cells on their ability to resist apoptosis onset and their production properties. The expression of mmu-miR-466h-5p in the engineered anti-miR-466h CHO cell line was significantly lower than in the negative control and the parental CHO cells. These engineered cells reached higher maximum viable cell density and extended viability compared with negative control and parental CHO cells in batch cell cultures which resulted in the 53.8% and 41.6% increase of integral viable cells. The extended viability of anti-miR-466h CHO cells was the result of delayed Caspase-3/7 activation by more than 35h, and the increased levels of its anti-apoptotic gene targets (smo, stat5a, dad1, birc6, and bcl2l2) to between 2.1- and 12.5-fold compared with the negative control CHO in apoptotic conditions. The expression of secreted alkaline phosphatase (SEAP) increased 43% and the cell-specific productivity increased 11% in the stable pools of anti-miR-466h CHO compared with the stable pools of negative control CHO cells. The above results demonstrate the potential of this novel approach to create more productive cell lines through stable manipulation of specific miRNA expression.

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  • Inhibition of the mmu-miR-466 h microRNA was shown to increase apoptosis resistance of these cells.

    “Also, as discussed earlier, inhibition of the mmu-miR-466 h microRNA was shown to increase apoptosis resistance of these cells (Druz et al., 2011;Druz et al., 2013).”

    From Therapeutic glycoprotein production in mammalian cells (2017) · cites this paper as Druz et al., 2013

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Cited by

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Authors
Druz, A., Son, Y., Betenbaugh, M., & Shiloach, J.
Affiliations
  • National Institutes of Health (US)
  • Johns Hopkins University (US)
Corresponding author
Shiloach, Joseph
ORCID
Venue
Metabolic Engineering
Publisher
Elsevier BV
Published
2013-03
Volume
16
Pages
87-94
ISSN
1096-7176 (ISSN-L)
DOI
10.1016/j.ymben.2012.12.004
PMC
PMC3644357
OpenAlex
W2014053766
Cited by
73
References
0
Citations by year
2026: 2 · 2025: 3 · 2023: 2 · 2022: 3 · 2021: 5 · 2020: 7 · 2019: 5 · 2018: 8 · 2017: 8 · 2016: 4 · 2015: 9 · 2014: 12 · 2013: 5
FWCI
4.48
Type
journal article
Language
en
Open access
Licence
www.elsevier.com/tdm/userlicense/1.0/ (tdm)
Locations
  • Metabolic Engineering (publishedVersion)
  • PubMed (publishedVersion)
  • Europe PMC (PubMed Central) (acceptedVersion)
  • PubMed Central (acceptedVersion)
Funders
  • National Institutes of Health
  • National Institute of Diabetes and Digestive and Kidney Diseases (ZIADK070010)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Viral Infectious Diseases and Gene Expression in Insects
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › MicroRNA in disease regulation
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Cell death mechanisms and regulation
Keywords
microRNA inhibition, apoptosis resistance, CHO cells, caspase-3/7 activation, secreted alkaline phosphatase, cell-specific productivity, cell viability, bioprocess scale-up
MeSH terms
Animals, Cell Survival, Cricetulus, Cricetinae, CHO Cells, Apoptosis, Gene Silencing, MicroRNAs, Inhibitor of Apoptosis Proteins, Caspase 3, Caspase 7

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