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Re‐programming CHO cell metabolism using miR‐23 tips the balance towards a highly productive phenotype
Author(s) -
Kelly Paul S.,
Breen Laura,
Gallagher Clair,
Kelly Shane,
Henry Michael,
Lao Nga T.,
Meleady Paula,
O'Gorman Donal,
Clynes Martin,
Barron Niall
Publication year - 2015
Publication title -
biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.144
H-Index - 84
eISSN - 1860-7314
pISSN - 1860-6768
DOI - 10.1002/biot.201500101
Subject(s) - oxidative phosphorylation , metabolism , microbiology and biotechnology , biology , glycolysis , citric acid cycle , mitochondrion , biochemistry , metabolic engineering , bioenergetics , phenotype , cell growth , flux balance analysis , gene
Abstract microRNA engineering of CHO cells has already proved successful in enhancing various industrially relevant phenotypes and producing various recombinant products. A single miRNA's ability to interact with multiple mRNA targets allows their regulatory capacity to extend to processes such as cellular metabolism. Various metabolic states have previously been associated with particular CHO cell phenotypes such as glycolytic or oxidative metabolism accommodating growth and productivity, respectively. miR‐23 has previously been demonstrated to play a role in glutamate metabolism resulting in enhanced oxidative phosphorylation through the TCA cycle. Re‐programming cellular bioenergetics through miR‐23 could tip the balance, forcing mammalian production cells to be more productive by favoring metabolic channelling into oxidative metabolism. CHO clones depleted of miR‐23 using a miR‐sponge decoy demonstrated an average ∼three‐fold enhanced specific productivity with no impact on cell growth. Using a cell respirometer, mitochondrial activity was found to be enhanced by ∼30% at Complex I and II of the electron transport system. Additionally, label‐free proteomic analysis uncovered various potential novel targets of miR‐23 including LE™1 and IDH1 , both implicated in oxidative metabolism and mitochondrial activity. These results demonstrate miRNA‐based engineering as a route to re‐programming cellular metabolism resulting in increased productivity, without affecting growth.

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