Three new shRNA expression vectors targeting the CYP3A4 coding sequence to inhibit its expression
RNA interference (RNAi) is useful for selective gene silencing. Cytochrome P450 3A4 (CYP3A4), which metabolizes approximately 50% of drugs in clinical use, plays an important role in drug metabolism. In this study, we aimed to develop a short hairpin RNA (shRNA) to modulate CYP3A4 expression. Three...
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| Format: | Article |
| Language: | English |
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Elsevier
2014-10-01
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| Series: | Acta Pharmaceutica Sinica B |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2211383514000859 |
| _version_ | 1811342770840797184 |
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| author | Siyun Xu Yongsheng Xiao Li Li Lushan Yu Huidi Jiang Aiming Yu Su Zeng |
| author_facet | Siyun Xu Yongsheng Xiao Li Li Lushan Yu Huidi Jiang Aiming Yu Su Zeng |
| author_sort | Siyun Xu |
| collection | DOAJ |
| description | RNA interference (RNAi) is useful for selective gene silencing. Cytochrome P450 3A4 (CYP3A4), which metabolizes approximately 50% of drugs in clinical use, plays an important role in drug metabolism. In this study, we aimed to develop a short hairpin RNA (shRNA) to modulate CYP3A4 expression. Three new shRNAs (S1, S2 and S3) were designed to target the coding sequence (CDS) of CYP3A4, cloned into a shRNA expression vector, and tested in different cells. The mixture of three shRNAs produced optimal reduction (55%) in CYP3A4 CDS-luciferase activity in both CHL and HEK293 cells. Endogenous CYP3A4 expression in HepG2 cells was decreased about 50% at both mRNA and protein level after transfection of the mixture of three shRNAs. In contrast, CYP3A5 gene expression was not altered by the shRNAs, supporting the selectivity of CYP3A4 shRNAs. In addition, HepG2 cells transfected with CYP3A4 shRNAs were less sensitive to Ginkgolic acids, whose toxic metabolites are produced by CYP3A4. These results demonstrate that vector-based shRNAs could modulate CYP3A4 expression in cells through their actions on CYP3A4 CDS, and CYP3A4 shRNAs may be utilized to define the role of CYP3A4 in drug metabolism and toxicity. |
| first_indexed | 2024-04-13T19:18:00Z |
| format | Article |
| id | doaj.art-65f08306363a4d63ac93010f1d4356bf |
| institution | Directory Open Access Journal |
| issn | 2211-3835 2211-3843 |
| language | English |
| last_indexed | 2024-04-13T19:18:00Z |
| publishDate | 2014-10-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Acta Pharmaceutica Sinica B |
| spelling | doaj.art-65f08306363a4d63ac93010f1d4356bf2022-12-22T02:33:38ZengElsevierActa Pharmaceutica Sinica B2211-38352211-38432014-10-014535035710.1016/j.apsb.2014.08.003Three new shRNA expression vectors targeting the CYP3A4 coding sequence to inhibit its expressionSiyun Xu0Yongsheng Xiao1Li Li2Lushan Yu3Huidi Jiang4Aiming Yu5Su Zeng6Department of Pharmaceutical Analysis and Drug Metabolism, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, ChinaDepartment of Pharmaceutical Analysis and Drug Metabolism, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, ChinaDepartment of Pharmaceutical Analysis and Drug Metabolism, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, ChinaDepartment of Pharmaceutical Analysis and Drug Metabolism, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, ChinaDepartment of Pharmaceutical Analysis and Drug Metabolism, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, ChinaDepartment of Biochemistry & Molecular Medicine, UC-Davis Medical Center, Sacramento, CA 95817, USADepartment of Pharmaceutical Analysis and Drug Metabolism, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, ChinaRNA interference (RNAi) is useful for selective gene silencing. Cytochrome P450 3A4 (CYP3A4), which metabolizes approximately 50% of drugs in clinical use, plays an important role in drug metabolism. In this study, we aimed to develop a short hairpin RNA (shRNA) to modulate CYP3A4 expression. Three new shRNAs (S1, S2 and S3) were designed to target the coding sequence (CDS) of CYP3A4, cloned into a shRNA expression vector, and tested in different cells. The mixture of three shRNAs produced optimal reduction (55%) in CYP3A4 CDS-luciferase activity in both CHL and HEK293 cells. Endogenous CYP3A4 expression in HepG2 cells was decreased about 50% at both mRNA and protein level after transfection of the mixture of three shRNAs. In contrast, CYP3A5 gene expression was not altered by the shRNAs, supporting the selectivity of CYP3A4 shRNAs. In addition, HepG2 cells transfected with CYP3A4 shRNAs were less sensitive to Ginkgolic acids, whose toxic metabolites are produced by CYP3A4. These results demonstrate that vector-based shRNAs could modulate CYP3A4 expression in cells through their actions on CYP3A4 CDS, and CYP3A4 shRNAs may be utilized to define the role of CYP3A4 in drug metabolism and toxicity.http://www.sciencedirect.com/science/article/pii/S2211383514000859RNAiCytochrome P450CYP3A4shRNAChemosensitivity |
| spellingShingle | Siyun Xu Yongsheng Xiao Li Li Lushan Yu Huidi Jiang Aiming Yu Su Zeng Three new shRNA expression vectors targeting the CYP3A4 coding sequence to inhibit its expression Acta Pharmaceutica Sinica B RNAi Cytochrome P450 CYP3A4 shRNA Chemosensitivity |
| title | Three new shRNA expression vectors targeting the CYP3A4 coding sequence to inhibit its expression |
| title_full | Three new shRNA expression vectors targeting the CYP3A4 coding sequence to inhibit its expression |
| title_fullStr | Three new shRNA expression vectors targeting the CYP3A4 coding sequence to inhibit its expression |
| title_full_unstemmed | Three new shRNA expression vectors targeting the CYP3A4 coding sequence to inhibit its expression |
| title_short | Three new shRNA expression vectors targeting the CYP3A4 coding sequence to inhibit its expression |
| title_sort | three new shrna expression vectors targeting the cyp3a4 coding sequence to inhibit its expression |
| topic | RNAi Cytochrome P450 CYP3A4 shRNA Chemosensitivity |
| url | http://www.sciencedirect.com/science/article/pii/S2211383514000859 |
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