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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...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4345305/ https://www.ncbi.nlm.nih.gov/pubmed/25602583 http://dx.doi.org/10.1038/mtna.2014.73 |
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author | Sun, Guihua Yeh, Spencer Yele Yuan, Christine Wan-Yin Chiu, Margaret Jean-Yih Yung, Bryan Shing Hei Yen, Yun |
author_facet | Sun, Guihua Yeh, Spencer Yele Yuan, Christine Wan-Yin Chiu, Margaret Jean-Yih Yung, Bryan Shing Hei Yen, Yun |
author_sort | Sun, Guihua |
collection | PubMed |
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. |
format | Online Article Text |
id | pubmed-4345305 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-43453052015-03-09 Molecular Properties, Functional Mechanisms, and Applications of Sliced siRNA Sun, Guihua Yeh, Spencer Yele Yuan, Christine Wan-Yin Chiu, Margaret Jean-Yih Yung, Bryan Shing Hei Yen, Yun Mol Ther Nucleic Acids Original Article 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. Nature Publishing Group 2015-01 2015-01-20 /pmc/articles/PMC4345305/ /pubmed/25602583 http://dx.doi.org/10.1038/mtna.2014.73 Text en Copyright © 2015 American Society of Gene & Cell Therapy http://creativecommons.org/licenses/by-nc-sa/3.0/ This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/3.0/ |
spellingShingle | Original Article Sun, Guihua Yeh, Spencer Yele Yuan, Christine Wan-Yin Chiu, Margaret Jean-Yih Yung, Bryan Shing Hei Yen, Yun Molecular Properties, Functional Mechanisms, and Applications of 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 | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4345305/ https://www.ncbi.nlm.nih.gov/pubmed/25602583 http://dx.doi.org/10.1038/mtna.2014.73 |
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