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Subphysiological temperature induces pervasive alternative splicing in Chinese hamster ovary cells.
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.
Afiliação
  • Tzani I; National Institute for Bioprocessing Research and Training, Dublin, Ireland.
  • Monger C; National Institute for Bioprocessing Research and Training, Dublin, Ireland.
  • Motheramgari K; National Institute for Bioprocessing Research and Training, Dublin, Ireland.
  • Gallagher C; National Institute for Cellular Biotechnology, Dublin City University Glasnevin, Dublin, Ireland.
  • Hagan R; National Institute for Cellular Biotechnology, Dublin City University Glasnevin, Dublin, Ireland.
  • Kelly P; National Institute for Bioprocessing Research and Training, Dublin, Ireland.
  • Costello A; School of Chemical and Bioprocess Engineering, University College Dublin, Dublin, Ireland.
  • Meiller J; National Institute for Cellular Biotechnology, Dublin City University Glasnevin, Dublin, Ireland.
  • Floris P; National Institute for Cellular Biotechnology, Dublin City University Glasnevin, Dublin, Ireland.
  • Zhang L; National Institute for Cellular Biotechnology, Dublin City University Glasnevin, Dublin, Ireland.
  • Clynes M; National Institute for Bioprocessing Research and Training, Dublin, Ireland.
  • Bones J; Bioprocess R&D, Pfizer Inc., Andover, Massachusetts.
  • Barron N; National Institute for Cellular Biotechnology, Dublin City University Glasnevin, Dublin, Ireland.
  • Clarke C; National Institute for Bioprocessing Research and Training, Dublin, Ireland.
Biotechnol Bioeng ; 117(8): 2489-2503, 2020 08.
Article em En | MEDLINE | ID: mdl-32346860
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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Texto completo: 1 Base de dados: MEDLINE Assunto principal: RNA Mensageiro / Processamento Alternativo / Transcriptoma Limite: Animals Idioma: En Ano de publicação: 2020 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: RNA Mensageiro / Processamento Alternativo / Transcriptoma Limite: Animals Idioma: En Ano de publicação: 2020 Tipo de documento: Article