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The EF‐1α promoter maintains high‐level transgene expression from episomal vectors in transfected CHO‐K1 cells
In our previous study, we demonstrated that episomal vectors based on the characteristic sequence of matrix attachment regions (MARs) and containing the cytomegalovirus (CMV) promoter allow transgenes to be maintained episomally in Chinese hamster ovary (CHO) cells. However, the transgene expression...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5661254/ https://www.ncbi.nlm.nih.gov/pubmed/28557288 http://dx.doi.org/10.1111/jcmm.13216 |
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author | Wang, Xiaoyin Xu, Zhongjie Tian, Zhengwei Zhang, Xi Xu, Danhua Li, Qin Zhang, Junhe Wang, Tianyun |
author_facet | Wang, Xiaoyin Xu, Zhongjie Tian, Zhengwei Zhang, Xi Xu, Danhua Li, Qin Zhang, Junhe Wang, Tianyun |
author_sort | Wang, Xiaoyin |
collection | PubMed |
description | In our previous study, we demonstrated that episomal vectors based on the characteristic sequence of matrix attachment regions (MARs) and containing the cytomegalovirus (CMV) promoter allow transgenes to be maintained episomally in Chinese hamster ovary (CHO) cells. However, the transgene expression was unstable and the number of copies was low. In this study, we focused on enhancers, various promoters and promoter variants that could improve the transgene expression stability, expression magnitude (level) and the copy number of a MAR‐based episomal vector in CHO‐K1 cells. In comparison with the CMV promoter, the eukaryotic translation elongation factor 1 α (EF‐1α, gene symbol EEF1A1) promoter increased the transfection efficiency, the transgene expression, the proportion of expression‐positive clones and the copy number of the episomal vector in long‐term culture. By contrast, no significant positive effects were observed with an enhancer, CMV promoter variants or CAG promoter in the episomal vector in long‐term culture. Moreover, the high‐expression clones harbouring the EF‐1α promoter tended to be more stable in long‐term culture, even in the absence of selection pressure. According to these findings, we concluded that the EF‐1α promoter is a potent regulatory sequence for episomal vectors because it maintains high transgene expression, transgene stability and copy number. These results provide valuable information on improvement of transgene stability and the copy number of episomal vectors. |
format | Online Article Text |
id | pubmed-5661254 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-56612542017-11-02 The EF‐1α promoter maintains high‐level transgene expression from episomal vectors in transfected CHO‐K1 cells Wang, Xiaoyin Xu, Zhongjie Tian, Zhengwei Zhang, Xi Xu, Danhua Li, Qin Zhang, Junhe Wang, Tianyun J Cell Mol Med Original Articles In our previous study, we demonstrated that episomal vectors based on the characteristic sequence of matrix attachment regions (MARs) and containing the cytomegalovirus (CMV) promoter allow transgenes to be maintained episomally in Chinese hamster ovary (CHO) cells. However, the transgene expression was unstable and the number of copies was low. In this study, we focused on enhancers, various promoters and promoter variants that could improve the transgene expression stability, expression magnitude (level) and the copy number of a MAR‐based episomal vector in CHO‐K1 cells. In comparison with the CMV promoter, the eukaryotic translation elongation factor 1 α (EF‐1α, gene symbol EEF1A1) promoter increased the transfection efficiency, the transgene expression, the proportion of expression‐positive clones and the copy number of the episomal vector in long‐term culture. By contrast, no significant positive effects were observed with an enhancer, CMV promoter variants or CAG promoter in the episomal vector in long‐term culture. Moreover, the high‐expression clones harbouring the EF‐1α promoter tended to be more stable in long‐term culture, even in the absence of selection pressure. According to these findings, we concluded that the EF‐1α promoter is a potent regulatory sequence for episomal vectors because it maintains high transgene expression, transgene stability and copy number. These results provide valuable information on improvement of transgene stability and the copy number of episomal vectors. John Wiley and Sons Inc. 2017-05-30 2017-11 /pmc/articles/PMC5661254/ /pubmed/28557288 http://dx.doi.org/10.1111/jcmm.13216 Text en © 2017 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine. This is an open access article under the terms of the Creative Commons Attribution (http://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Original Articles Wang, Xiaoyin Xu, Zhongjie Tian, Zhengwei Zhang, Xi Xu, Danhua Li, Qin Zhang, Junhe Wang, Tianyun The EF‐1α promoter maintains high‐level transgene expression from episomal vectors in transfected CHO‐K1 cells |
title | The EF‐1α promoter maintains high‐level transgene expression from episomal vectors in transfected CHO‐K1 cells |
title_full | The EF‐1α promoter maintains high‐level transgene expression from episomal vectors in transfected CHO‐K1 cells |
title_fullStr | The EF‐1α promoter maintains high‐level transgene expression from episomal vectors in transfected CHO‐K1 cells |
title_full_unstemmed | The EF‐1α promoter maintains high‐level transgene expression from episomal vectors in transfected CHO‐K1 cells |
title_short | The EF‐1α promoter maintains high‐level transgene expression from episomal vectors in transfected CHO‐K1 cells |
title_sort | ef‐1α promoter maintains high‐level transgene expression from episomal vectors in transfected cho‐k1 cells |
topic | Original Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5661254/ https://www.ncbi.nlm.nih.gov/pubmed/28557288 http://dx.doi.org/10.1111/jcmm.13216 |
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