Optimization of the intron sequences combined with the CMV promoter increases recombinant protein expression in CHO cells
Abstract To meet the requirements of the biopharmaceutical industry, improving the yield of recombination therapeutic protein (RTP) from Chinese hamster ovary (CHO) cells is necessary. The human cytomegalovirus (CMV) promoter is widely used for RTP expression in CHO cells. To further improve RTP pro...
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Nature Portfolio
2025-01-01
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| Online Access: | https://doi.org/10.1038/s41598-025-87941-5 |
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| author | Ting-Ting Xu Yan-Fang Wang Jing-Jia Yuan Chun-Liu Mi Shao-Lei Geng Xiao-yin Wang Tian-Yun Wang |
| author_facet | Ting-Ting Xu Yan-Fang Wang Jing-Jia Yuan Chun-Liu Mi Shao-Lei Geng Xiao-yin Wang Tian-Yun Wang |
| author_sort | Ting-Ting Xu |
| collection | DOAJ |
| description | Abstract To meet the requirements of the biopharmaceutical industry, improving the yield of recombination therapeutic protein (RTP) from Chinese hamster ovary (CHO) cells is necessary. The human cytomegalovirus (CMV) promoter is widely used for RTP expression in CHO cells. To further improve RTP production, we truncated the human CMV intron and further evaluated the effect of four synthetic introns, including ctEF-1α first, EF-1α first, chimeric, and β-globin introns combined with the CMV promoter on recombinant expression levels in transient and stably recombinant CHO cells. The results showed that the truncated, EF-1α first, chimeric, and β-globin introns can significantly improve stable transgene expression in CHO cells. The qPCR results indicated that the mRNA level of transgene increased through optimizing intron sequences combined with the CMV promoter. Transcriptomics analysis was performed and found that differential expression of genes involved in mRNA processing, RNA export from nucleus, cytoplasmic translation, transcriptional activation and cell cycle regulation. In conclusion, optimization of the intron sequences combined with the CMV promoter can achieve a higher yield of recombinant proteins in CHO cells. This will be valuable for generating CHO cell lines with high productivity for industrial applications. |
| format | Article |
| id | doaj-art-993ba21e2ff2415a827f455b6f864ad2 |
| institution | Kabale University |
| issn | 2045-2322 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj-art-993ba21e2ff2415a827f455b6f864ad22025-02-02T12:15:40ZengNature PortfolioScientific Reports2045-23222025-01-0115111110.1038/s41598-025-87941-5Optimization of the intron sequences combined with the CMV promoter increases recombinant protein expression in CHO cellsTing-Ting Xu0Yan-Fang Wang1Jing-Jia Yuan2Chun-Liu Mi3Shao-Lei Geng4Xiao-yin Wang5Tian-Yun Wang6International Joint Research Laboratory for Recombinant Pharmaceutical Protein Expression System of Henan, Xinxiang Medical UniversityThe First Affiliated Hospital, Xinxiang Medical UniversityInternational Joint Research Laboratory for Recombinant Pharmaceutical Protein Expression System of Henan, Xinxiang Medical UniversityInternational Joint Research Laboratory for Recombinant Pharmaceutical Protein Expression System of Henan, Xinxiang Medical UniversityInternational Joint Research Laboratory for Recombinant Pharmaceutical Protein Expression System of Henan, Xinxiang Medical UniversityInternational Joint Research Laboratory for Recombinant Pharmaceutical Protein Expression System of Henan, Xinxiang Medical UniversityInternational Joint Research Laboratory for Recombinant Pharmaceutical Protein Expression System of Henan, Xinxiang Medical UniversityAbstract To meet the requirements of the biopharmaceutical industry, improving the yield of recombination therapeutic protein (RTP) from Chinese hamster ovary (CHO) cells is necessary. The human cytomegalovirus (CMV) promoter is widely used for RTP expression in CHO cells. To further improve RTP production, we truncated the human CMV intron and further evaluated the effect of four synthetic introns, including ctEF-1α first, EF-1α first, chimeric, and β-globin introns combined with the CMV promoter on recombinant expression levels in transient and stably recombinant CHO cells. The results showed that the truncated, EF-1α first, chimeric, and β-globin introns can significantly improve stable transgene expression in CHO cells. The qPCR results indicated that the mRNA level of transgene increased through optimizing intron sequences combined with the CMV promoter. Transcriptomics analysis was performed and found that differential expression of genes involved in mRNA processing, RNA export from nucleus, cytoplasmic translation, transcriptional activation and cell cycle regulation. In conclusion, optimization of the intron sequences combined with the CMV promoter can achieve a higher yield of recombinant proteins in CHO cells. This will be valuable for generating CHO cell lines with high productivity for industrial applications.https://doi.org/10.1038/s41598-025-87941-5Recombinant therapeutic proteinChinese hamster ovary cellsExpression vectorPromoterIntron |
| spellingShingle | Ting-Ting Xu Yan-Fang Wang Jing-Jia Yuan Chun-Liu Mi Shao-Lei Geng Xiao-yin Wang Tian-Yun Wang Optimization of the intron sequences combined with the CMV promoter increases recombinant protein expression in CHO cells Scientific Reports Recombinant therapeutic protein Chinese hamster ovary cells Expression vector Promoter Intron |
| title | Optimization of the intron sequences combined with the CMV promoter increases recombinant protein expression in CHO cells |
| title_full | Optimization of the intron sequences combined with the CMV promoter increases recombinant protein expression in CHO cells |
| title_fullStr | Optimization of the intron sequences combined with the CMV promoter increases recombinant protein expression in CHO cells |
| title_full_unstemmed | Optimization of the intron sequences combined with the CMV promoter increases recombinant protein expression in CHO cells |
| title_short | Optimization of the intron sequences combined with the CMV promoter increases recombinant protein expression in CHO cells |
| title_sort | optimization of the intron sequences combined with the cmv promoter increases recombinant protein expression in cho cells |
| topic | Recombinant therapeutic protein Chinese hamster ovary cells Expression vector Promoter Intron |
| url | https://doi.org/10.1038/s41598-025-87941-5 |
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