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CMV promoter mutants with a reduced propensity to productivity loss in CHO cells

The major immediate-early promoter and enhancer of the human cytomegalovirus (hCMV-MIE) is one of the most potent DNA elements driving recombinant gene expression in mammalian cells. Therefore, it is widely employed not only in research but also in large-scale industrial applications, e.g. for the p...

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Autores principales: Moritz, Benjamin, Becker, Peter B., Göpfert, Ulrich
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4652263/
https://www.ncbi.nlm.nih.gov/pubmed/26581326
http://dx.doi.org/10.1038/srep16952
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author Moritz, Benjamin
Becker, Peter B.
Göpfert, Ulrich
author_facet Moritz, Benjamin
Becker, Peter B.
Göpfert, Ulrich
author_sort Moritz, Benjamin
collection PubMed
description The major immediate-early promoter and enhancer of the human cytomegalovirus (hCMV-MIE) is one of the most potent DNA elements driving recombinant gene expression in mammalian cells. Therefore, it is widely employed not only in research but also in large-scale industrial applications, e.g. for the production of therapeutic antibodies in Chinese hamster ovary cells (CHO). As we have reported previously, multi-site methylation of hCMV-MIE is linked to productivity loss in permanently transfected CHO cells lines. In particular, the cytosine located 179 bp upstream of the transcription start site (C-179) is frequently methylated. Therefore, our objective was to study whether mutation of C-179 and other cytosines within hCMV-MIE might lessen the instability of transgene expression. We discovered that the single mutation of C-179 to G can significantly stabilise the production of recombinant protein under control of hCMV-MIE in permanently transfected CHO cells.
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spelling pubmed-46522632015-11-24 CMV promoter mutants with a reduced propensity to productivity loss in CHO cells Moritz, Benjamin Becker, Peter B. Göpfert, Ulrich Sci Rep Article The major immediate-early promoter and enhancer of the human cytomegalovirus (hCMV-MIE) is one of the most potent DNA elements driving recombinant gene expression in mammalian cells. Therefore, it is widely employed not only in research but also in large-scale industrial applications, e.g. for the production of therapeutic antibodies in Chinese hamster ovary cells (CHO). As we have reported previously, multi-site methylation of hCMV-MIE is linked to productivity loss in permanently transfected CHO cells lines. In particular, the cytosine located 179 bp upstream of the transcription start site (C-179) is frequently methylated. Therefore, our objective was to study whether mutation of C-179 and other cytosines within hCMV-MIE might lessen the instability of transgene expression. We discovered that the single mutation of C-179 to G can significantly stabilise the production of recombinant protein under control of hCMV-MIE in permanently transfected CHO cells. Nature Publishing Group 2015-11-19 /pmc/articles/PMC4652263/ /pubmed/26581326 http://dx.doi.org/10.1038/srep16952 Text en Copyright © 2015, Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Moritz, Benjamin
Becker, Peter B.
Göpfert, Ulrich
CMV promoter mutants with a reduced propensity to productivity loss in CHO cells
title CMV promoter mutants with a reduced propensity to productivity loss in CHO cells
title_full CMV promoter mutants with a reduced propensity to productivity loss in CHO cells
title_fullStr CMV promoter mutants with a reduced propensity to productivity loss in CHO cells
title_full_unstemmed CMV promoter mutants with a reduced propensity to productivity loss in CHO cells
title_short CMV promoter mutants with a reduced propensity to productivity loss in CHO cells
title_sort cmv promoter mutants with a reduced propensity to productivity loss in cho cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4652263/
https://www.ncbi.nlm.nih.gov/pubmed/26581326
http://dx.doi.org/10.1038/srep16952
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