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...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2022-05-01
|
| Series: | Frontiers in Bioengineering and Biotechnology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fbioe.2022.840600/full |
| _version_ | 1811239599806087168 |
|---|---|
| author | Qin Li Qin Li Rui-Fang Yan Yong-Xiao Yang Chun-liu Mi Chun-liu Mi Yan-long Jia Tian-Yun Wang Tian-Yun Wang |
| author_facet | Qin Li Qin Li Rui-Fang Yan Yong-Xiao Yang Chun-liu Mi Chun-liu Mi Yan-long Jia Tian-Yun Wang Tian-Yun Wang |
| author_sort | Qin Li |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-04-12T13:03:49Z |
| format | Article |
| id | doaj.art-b0e1dbd268e74ecead489c9366e7d334 |
| institution | Directory Open Access Journal |
| issn | 2296-4185 |
| language | English |
| last_indexed | 2024-04-12T13:03:49Z |
| publishDate | 2022-05-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Bioengineering and Biotechnology |
| spelling | doaj.art-b0e1dbd268e74ecead489c9366e7d3342022-12-22T03:32:06ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852022-05-011010.3389/fbioe.2022.840600840600Stabilizing and Anti-Repressor Elements Effectively Increases Transgene Expression in Transfected CHO CellsQin Li0Qin Li1Rui-Fang Yan2Yong-Xiao Yang3Chun-liu Mi4Chun-liu Mi5Yan-long Jia6Tian-Yun Wang7Tian-Yun Wang8School of Basic Medicine, Xinxiang Medical University, Xinxiang, ChinaInternational Joint Research Laboratory for Recombinant Pharmaceutical Protein Expression System of Henan, Xinxiang Medical University, Xinxiang, ChinaSchool of Basic Medicine, Xinxiang Medical University, Xinxiang, ChinaInternational Joint Research Laboratory for Recombinant Pharmaceutical Protein Expression System of Henan, Xinxiang Medical University, Xinxiang, ChinaSchool of Basic Medicine, Xinxiang Medical University, Xinxiang, ChinaInternational Joint Research Laboratory for Recombinant Pharmaceutical Protein Expression System of Henan, Xinxiang Medical University, Xinxiang, ChinaInternational Joint Research Laboratory for Recombinant Pharmaceutical Protein Expression System of Henan, Xinxiang Medical University, Xinxiang, ChinaSchool of Basic Medicine, Xinxiang Medical University, Xinxiang, ChinaInternational Joint Research Laboratory for Recombinant Pharmaceutical Protein Expression System of Henan, Xinxiang Medical University, Xinxiang, ChinaChinese 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.https://www.frontiersin.org/articles/10.3389/fbioe.2022.840600/fullChinese hamster ovary cellstabilizing and anti-repressor elementstransgene expressionepigenetic regulatory elementsgreen fluorescent protein |
| spellingShingle | Qin Li Qin Li Rui-Fang Yan Yong-Xiao Yang Chun-liu Mi Chun-liu Mi Yan-long Jia Tian-Yun Wang Tian-Yun Wang Stabilizing and Anti-Repressor Elements Effectively Increases Transgene Expression in Transfected CHO Cells Frontiers in Bioengineering and Biotechnology Chinese hamster ovary cell stabilizing and anti-repressor elements transgene expression epigenetic regulatory elements green fluorescent protein |
| 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 | Chinese hamster ovary cell stabilizing and anti-repressor elements transgene expression epigenetic regulatory elements green fluorescent protein |
| url | https://www.frontiersin.org/articles/10.3389/fbioe.2022.840600/full |
| work_keys_str_mv | AT qinli stabilizingandantirepressorelementseffectivelyincreasestransgeneexpressionintransfectedchocells AT qinli stabilizingandantirepressorelementseffectivelyincreasestransgeneexpressionintransfectedchocells AT ruifangyan stabilizingandantirepressorelementseffectivelyincreasestransgeneexpressionintransfectedchocells AT yongxiaoyang stabilizingandantirepressorelementseffectivelyincreasestransgeneexpressionintransfectedchocells AT chunliumi stabilizingandantirepressorelementseffectivelyincreasestransgeneexpressionintransfectedchocells AT chunliumi stabilizingandantirepressorelementseffectivelyincreasestransgeneexpressionintransfectedchocells AT yanlongjia stabilizingandantirepressorelementseffectivelyincreasestransgeneexpressionintransfectedchocells AT tianyunwang stabilizingandantirepressorelementseffectivelyincreasestransgeneexpressionintransfectedchocells AT tianyunwang stabilizingandantirepressorelementseffectivelyincreasestransgeneexpressionintransfectedchocells |