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Stabilizing and Anti-Repressor Elements Effectively Increases Transgene Expression in Transfected CHO Cells

Chinese hamster ovary (CHO) cells are currently the most widely used host cells for recombinant therapeutic protein (RTP) production. Currently, the RTP yields need to increase further to meet the market needs and reduce costs. In this study, three stabilizing and anti-repressor (SAR) elements from...

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Autores principales: Li, Qin, Yan, Rui-Fang, Yang, Yong-Xiao, Mi, Chun-liu, Jia, Yan-long, Wang, Tian-Yun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9199445/
https://www.ncbi.nlm.nih.gov/pubmed/35721852
http://dx.doi.org/10.3389/fbioe.2022.840600
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author Li, Qin
Yan, Rui-Fang
Yang, Yong-Xiao
Mi, Chun-liu
Jia, Yan-long
Wang, Tian-Yun
author_facet Li, Qin
Yan, Rui-Fang
Yang, Yong-Xiao
Mi, Chun-liu
Jia, Yan-long
Wang, Tian-Yun
author_sort Li, Qin
collection PubMed
description Chinese hamster ovary (CHO) cells are currently the most widely used host cells for recombinant therapeutic protein (RTP) production. Currently, the RTP yields need to increase further to meet the market needs and reduce costs. In this study, three stabilizing and anti-repressor (SAR) elements from the human genome were selected, including human SAR7, SAR40, and SAR44 elements. SAR elements were cloned upstream of the promoter in the eukaryotic vector, followed by transfection into CHO cells, and were screened under G418 pressure. Flow cytometry was used to detect enhanced green fluorescent protein (eGFP) expression levels. The gene copy numbers and mRNA expression levels were determined through quantitative real-time PCR. Furthermore, the effect of the stronger SAR elements on adalimumab was investigated. The results showed that transgene expression levels in the SAR-containing vectors were higher than that of the control vector, and SAR7 and SAR40 significantly increased and maintained the long-term expression of the transgene in CHO cells. In addition, the transgene expression level increase was related with gene copy numbers and mRNA expression levels. Collectively, SAR elements can enhance the transgene expression and maintain the long-term expression of a transgene in transfected CHO cells, which may be used to increase recombinant protein production in CHO cells.
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spelling pubmed-91994452022-06-16 Stabilizing and Anti-Repressor Elements Effectively Increases Transgene Expression in Transfected CHO Cells Li, Qin Yan, Rui-Fang Yang, Yong-Xiao Mi, Chun-liu Jia, Yan-long Wang, Tian-Yun Front Bioeng Biotechnol Bioengineering and Biotechnology Chinese hamster ovary (CHO) cells are currently the most widely used host cells for recombinant therapeutic protein (RTP) production. Currently, the RTP yields need to increase further to meet the market needs and reduce costs. In this study, three stabilizing and anti-repressor (SAR) elements from the human genome were selected, including human SAR7, SAR40, and SAR44 elements. SAR elements were cloned upstream of the promoter in the eukaryotic vector, followed by transfection into CHO cells, and were screened under G418 pressure. Flow cytometry was used to detect enhanced green fluorescent protein (eGFP) expression levels. The gene copy numbers and mRNA expression levels were determined through quantitative real-time PCR. Furthermore, the effect of the stronger SAR elements on adalimumab was investigated. The results showed that transgene expression levels in the SAR-containing vectors were higher than that of the control vector, and SAR7 and SAR40 significantly increased and maintained the long-term expression of the transgene in CHO cells. In addition, the transgene expression level increase was related with gene copy numbers and mRNA expression levels. Collectively, SAR elements can enhance the transgene expression and maintain the long-term expression of a transgene in transfected CHO cells, which may be used to increase recombinant protein production in CHO cells. Frontiers Media S.A. 2022-05-26 /pmc/articles/PMC9199445/ /pubmed/35721852 http://dx.doi.org/10.3389/fbioe.2022.840600 Text en Copyright © 2022 Li, Yan, Yang, Mi, Jia and Wang. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Bioengineering and Biotechnology
Li, Qin
Yan, Rui-Fang
Yang, Yong-Xiao
Mi, Chun-liu
Jia, Yan-long
Wang, Tian-Yun
Stabilizing and Anti-Repressor Elements Effectively Increases Transgene Expression in Transfected CHO Cells
title Stabilizing and Anti-Repressor Elements Effectively Increases Transgene Expression in Transfected CHO Cells
title_full Stabilizing and Anti-Repressor Elements Effectively Increases Transgene Expression in Transfected CHO Cells
title_fullStr Stabilizing and Anti-Repressor Elements Effectively Increases Transgene Expression in Transfected CHO Cells
title_full_unstemmed Stabilizing and Anti-Repressor Elements Effectively Increases Transgene Expression in Transfected CHO Cells
title_short Stabilizing and Anti-Repressor Elements Effectively Increases Transgene Expression in Transfected CHO Cells
title_sort stabilizing and anti-repressor elements effectively increases transgene expression in transfected cho cells
topic Bioengineering and Biotechnology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9199445/
https://www.ncbi.nlm.nih.gov/pubmed/35721852
http://dx.doi.org/10.3389/fbioe.2022.840600
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