中国仓鼠卵巢细胞
RNA剪接
选择性拼接
生物
转录组
外显子
基因
细胞生物学
信使核糖核酸
基因表达
外显子跳跃
分子生物学
RNA序列
核糖核酸
细胞培养
遗传学
作者
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
摘要
Abstract 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.
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