Type

Journal Article

Authors

Niall Barron
Martin Clynes
Colin Clarke
Nga T Lao
Clair Gallagher
Paul Kelly
Noelia Sanchez

Subjects

Biochemistry

Topics
genetics cricetinae genetic vectors antagonists inhibitors mirn7 microrna mouse antibodies monoclonal biosynthesis bioreactors cricetulus metabolism longevity cho cells micrornas animals cell culture techniques recombinant proteins cell count

CHO cell culture longevity and recombinant protein yield are enhanced by depletion of miR-7 activity via sponge decoy vectors. (2013)

Abstract Improving the efficiency of recombinant protein production by CHO cells is highly desirable as more complex proteins (MAbs, fusion proteins, blood/clotting factors, etc.) go into development and come onto the market. Previous reports have shown that microRNA (miRNA)-7 overexpression arrests the growth of CHO cells and that its depletion increases the proliferation of various cell types. In this study we generated stable CHO clones that overexpressed a miR-7-specific decoy transcript (sponge) downstream of a green fluorescent protein reporter gene. The miR-7 sponge efficiently diverted miR-7 away from its endogenous targets as exemplified by the increased expression of CDC7. Although the sponge effectively sequestered miR-7, it also appeared to protect the bound miRNA sequence from degradation in the cell, as exemplified by the apparent increase in mature miR-7 levels without any change in primary transcription. Phenotypically, CHO clones with sequestered miR-7 displayed improved maximum cell density (40%), significantly improved viability and an almost two-fold increase in yield of secreted protein in a fed-batch culture. These findings demonstrate that miRNA sponge transcripts could potentially be used in cell line development projects to generate producer clones that grow to higher densities and last longer in the bioreactor - thereby improving product yield.
Collections Ireland -> Dublin City University -> PubMed

Full list of authors on original publication

Niall Barron, Martin Clynes, Colin Clarke, Nga T Lao, Clair Gallagher, Paul Kelly, Noelia Sanchez

Experts in our system

1
Niall Barron
Dublin City University
Total Publications: 44
 
2
Martin Clynes
Dublin City University
Total Publications: 209
 
3
Colin Clarke
Dublin City University
Total Publications: 26
 
4
Nga T Lao
Dublin City University
Total Publications: 7
 
5
Clair Gallagher
Dublin City University
Total Publications: 12
 
6
Paul S. Kelly
Dublin City University
Total Publications: 11