Mechanisms underlying epigenetic and transcriptional heterogeneity in Chinese hamster ovary (CHO) cell lines

BMC Biotechnology
Nathalie VeithStella Marie Reamon-Buettner

Abstract

Recombinant cell lines developed for therapeutic antibody production often suffer instability or lose recombinant protein expression during long-term culture. Heterogeneous gene expression among cell line subclones may result from epigenetic modifications of DNA or histones, the protein component of chromatin. We thus investigated in such cell lines, DNA methylation and the chromatin environment along the human eukaryotic translation elongation factor 1 alpha 1 (EEF1A1) promoter in an antibody protein-expression vector which was integrated into the Chinese hamster ovary (CHO) cell line genome. We analyzed four PT1-CHO cell lines which exhibited losses of protein expression at advanced passage number (>P35) growing in adherent conditions and in culture medium with 10 % FCS. These cell lines exhibited different integration sites and transgene copy numbers as determined by fluorescence in situ hybridization (FISH) and quantitative PCR (qPCR), respectively. By qRT-PCR, we analyzed the recombinant mRNA expression and correlated it with DNA methylation and with results from various approaches interrogating the chromatin landscape along the EEF1A1 promoter region. Each PT1-CHO cell line displayed specific epigenetic signatures or chro...Continue Reading

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Citations

Sep 13, 2016·Biotechnology and Bioengineering·Anett RitterHolger Laux
Sep 30, 2016·Applied Microbiology and Biotechnology·Lilian Rumi TsurutaAna Maria Moro
Apr 2, 2017·Journal of Biotechnology·B MüllerR Hoffrogge
Aug 26, 2018·Biotechnology and Bioengineering·Arpan A BandyopadhyayWei-Shou Hu
Apr 19, 2019·Biotechnology and Bioengineering·Vania BertrandMassimo Morbidelli
May 7, 2020·Applied Microbiology and Biotechnology·Tian-Yun Wang, Xiao Guo
Jun 25, 2020·Current Research in Biotechnology·Hussain DahodwalaSusan T Sharfstein

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Methods Mentioned

BETA
ubiquitination
PCR
FCS
footprinting
immunoprecipitation
ChIP
acetylation
fluorescence microscopy

Software Mentioned

GraphPad Prism
CLC Workbench
NuPoP

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