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Subphysiological temperature induces pervasive alternative splicing in Chinese hamster ovary cells
Author(s) -
Tzani Ioanna,
Monger Craig,
Motheramgari Krishna,
Gallagher Clair,
Hagan Ryan,
Kelly Paul,
Costello Alan,
Meiller Justine,
Floris Patrick,
Zhang Lin,
Clynes Martin,
Bones Jonathan,
Barron Niall,
Clarke Colin
Publication year - 2020
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.27365
Subject(s) - chinese hamster ovary cell , rna splicing , alternative splicing , biology , transcriptome , exon , gene , microbiology and biotechnology , messenger rna , gene expression , exon skipping , rna seq , rna , cell culture , genetics
RNA sequencing (RNASeq) has been widely used to associate alterations in Chinese hamster ovary (CHO) cell gene expression with bioprocess phenotypes; however, alternative messenger RNA (mRNA) splicing, has thus far, received little attention. In this study, we utilized RNASeq for transcriptomic analysis of a monoclonal antibody (mAb) producing CHO K1 cell line subjected to a temperature shift. More than 2,465 instances of differential splicing were observed 24 hr after the reduction of cell culture temperature. A total of 1,197 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 quantitative polymerase chain reaction in the mAb‐producing CHO K1 cell line used for RNASeq and a further two CHO K1 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 these changes in mRNA sequence on cellular phenotype, this study demonstrates that alternative splicing analysis can be utilized to gain a deeper understanding of post‐transcriptional regulation in CHO cells during biopharmaceutical production.