Molecular Properties, Functional Mechanisms, and Applications of Sliced siRNA
Using pre-miR-451 as a model molecule, we have characterized the general molecular properties of small hairpin RNAs that are processed into potent small interfering RNAs (siRNA) by Argonaute2 (Ago2). The Ago2-sliced siRNAs (sli-siRNAs) have the same silencing potency as the classical Dicer diced siR...
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| Format: | Article |
| Language: | English |
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Elsevier
2015-01-01
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| Series: | Molecular Therapy: Nucleic Acids |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2162253116300051 |
| _version_ | 1818544348235563008 |
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| author | Guihua Sun Spencer Yele Yeh Christine Wan-Yin Yuan Margaret Jean-Yih Chiu Bryan Shing Hei Yung Yun Yen |
| author_facet | Guihua Sun Spencer Yele Yeh Christine Wan-Yin Yuan Margaret Jean-Yih Chiu Bryan Shing Hei Yung Yun Yen |
| author_sort | Guihua Sun |
| collection | DOAJ |
| description | Using pre-miR-451 as a model molecule, we have characterized the general molecular properties of small hairpin RNAs that are processed into potent small interfering RNAs (siRNA) by Argonaute2 (Ago2). The Ago2-sliced siRNAs (sli-siRNAs) have the same silencing potency as the classical Dicer diced siRNAs (di-siRNAs) but have dramatically reduced unwanted sense strand activities. We have built vectors with the constitutive or inducible U6 promoter that can express sli-siRNAs in mammalian cells, in which the sli-siRNAs can be correctly processed to repress target genes. As a proof of principle for potential applications of sli-siRNAs in vivo, we show that the expression of one Ago2 shRNA-1148 in HCT-116 colon cancer cells knocked down RRM2 expression and reduced the proliferation and invasiveness of the cells. The defined sli-siRNA model molecules and the expression systems established in this study will facilitate the design and application of sli-siRNAs as novel potent RNAi triggers with reduced off-target effects. |
| first_indexed | 2024-12-11T22:47:19Z |
| format | Article |
| id | doaj.art-586052314f834647bd79a3e850b44f73 |
| institution | Directory Open Access Journal |
| issn | 2162-2531 |
| language | English |
| last_indexed | 2024-12-11T22:47:19Z |
| publishDate | 2015-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Nucleic Acids |
| spelling | doaj.art-586052314f834647bd79a3e850b44f732022-12-22T00:47:34ZengElsevierMolecular Therapy: Nucleic Acids2162-25312015-01-014C10.1038/mtna.2014.73Molecular Properties, Functional Mechanisms, and Applications of Sliced siRNAGuihua Sun0Spencer Yele Yeh1Christine Wan-Yin Yuan2Margaret Jean-Yih Chiu3Bryan Shing Hei Yung4Yun Yen5Department of Molecular Pharmacology, Beckman Research Institute of the City of Hope, Duarte, California, USASummer Interns, Beckman Research Institute of the City of Hope, Duarte, California, USASummer Interns, Beckman Research Institute of the City of Hope, Duarte, California, USASummer Interns, Beckman Research Institute of the City of Hope, Duarte, California, USASummer Interns, Beckman Research Institute of the City of Hope, Duarte, California, USADepartment of Molecular Pharmacology, Beckman Research Institute of the City of Hope, Duarte, California, USAUsing pre-miR-451 as a model molecule, we have characterized the general molecular properties of small hairpin RNAs that are processed into potent small interfering RNAs (siRNA) by Argonaute2 (Ago2). The Ago2-sliced siRNAs (sli-siRNAs) have the same silencing potency as the classical Dicer diced siRNAs (di-siRNAs) but have dramatically reduced unwanted sense strand activities. We have built vectors with the constitutive or inducible U6 promoter that can express sli-siRNAs in mammalian cells, in which the sli-siRNAs can be correctly processed to repress target genes. As a proof of principle for potential applications of sli-siRNAs in vivo, we show that the expression of one Ago2 shRNA-1148 in HCT-116 colon cancer cells knocked down RRM2 expression and reduced the proliferation and invasiveness of the cells. The defined sli-siRNA model molecules and the expression systems established in this study will facilitate the design and application of sli-siRNAs as novel potent RNAi triggers with reduced off-target effects.http://www.sciencedirect.com/science/article/pii/S2162253116300051Ago2agshRNAagsiRNARNAisiRNAsliced siRNA |
| spellingShingle | Guihua Sun Spencer Yele Yeh Christine Wan-Yin Yuan Margaret Jean-Yih Chiu Bryan Shing Hei Yung Yun Yen Molecular Properties, Functional Mechanisms, and Applications of Sliced siRNA Molecular Therapy: Nucleic Acids Ago2 agshRNA agsiRNA RNAi siRNA sliced siRNA |
| title | Molecular Properties, Functional Mechanisms, and Applications of Sliced siRNA |
| title_full | Molecular Properties, Functional Mechanisms, and Applications of Sliced siRNA |
| title_fullStr | Molecular Properties, Functional Mechanisms, and Applications of Sliced siRNA |
| title_full_unstemmed | Molecular Properties, Functional Mechanisms, and Applications of Sliced siRNA |
| title_short | Molecular Properties, Functional Mechanisms, and Applications of Sliced siRNA |
| title_sort | molecular properties functional mechanisms and applications of sliced sirna |
| topic | Ago2 agshRNA agsiRNA RNAi siRNA sliced siRNA |
| url | http://www.sciencedirect.com/science/article/pii/S2162253116300051 |
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