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Sub physiological temperature induces pervasive alternative splicing in Chinese hamster ovary cells

By Ioanna Tzani, Craig Monger, Krishna Motheramgari, Clair Gallagher, Ryan Hagan, Paul Kelly, Alan Costello, Justine Meiller, Patrick Floris, Lin Zhang, Martin Clynes, Jonathan Bones, Niall Barron, Colin Clarke

Posted 03 Dec 2019
bioRxiv DOI: 10.1101/863175

RNA sequencing (RNASeq) has been widely used to associate alterations in Chinese hamster ovary (CHO) cell gene expression with bioprocess phenotypes, however alternative mRNA splicing, has thus far, received little attention. In this study, we utilised RNASeq for transcriptomic analysis of a monoclonal antibody producing CHOK1 cell line subjected to a temperature shift. More than 2,365 instances of differential splicing were observed 24hrs after the reduction of cell culture temperature. 1,163 of these alternative splicing events were identified in genes where no changes in abundance were detected by standard differential expression analysis. Ten examples of alternative splicing were selected for independent validation using qPCR in the monoclonal antibody producing CHOK1 line used for RNASeq and a further 2 CHOK1 cell lines. This analysis provided evidence that exon skipping and mutually exclusive splicing events occur in genes linked to the cellular response to changes in temperature and mitochondrial function. While further work is required to determine the impact of changes in mRNA sequence on cellular phenotype, this study demonstrates that alternative splicing analysis can be utilised to gain a deeper understanding of post-transcriptional regulation in CHO cells during biopharmaceutical production.

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