Both strands of siRNA have potential to guide posttranscriptional gene silencing in mammalian cells.

Despite the widespread application of RNA interference (RNAi) as a research tool for diverse purposes, the key step of strand selection of siRNAs during the formation of RNA-induced silencing complex (RISC) remains poorly understood. Here, using siRNAs targeted to the complementary region of Survivi...

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Main Authors: Jun-Xia Wei, Jie Yang, Ji-Feng Sun, Lin-Tao Jia, Yong Zhang, Hui-Zhong Zhang, Xia Li, Yan-Ling Meng, Li-Bo Yao, An-Gang Yang
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2009-01-01
Series:PLoS ONE
Online Access:https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0005382&type=printable
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author Jun-Xia Wei
Jie Yang
Ji-Feng Sun
Lin-Tao Jia
Yong Zhang
Hui-Zhong Zhang
Xia Li
Yan-Ling Meng
Li-Bo Yao
An-Gang Yang
author_facet Jun-Xia Wei
Jie Yang
Ji-Feng Sun
Lin-Tao Jia
Yong Zhang
Hui-Zhong Zhang
Xia Li
Yan-Ling Meng
Li-Bo Yao
An-Gang Yang
author_sort Jun-Xia Wei
collection DOAJ
description Despite the widespread application of RNA interference (RNAi) as a research tool for diverse purposes, the key step of strand selection of siRNAs during the formation of RNA-induced silencing complex (RISC) remains poorly understood. Here, using siRNAs targeted to the complementary region of Survivin and the effector protease receptor 1 (EPR-1), we show that both strands of the siRNA duplex can find their target mRNA and are equally eligible for assembly into Argonaute 2 (Ago2) of RISC in HEK293 cells. Transfection of the synthetic siRNA duplexes with different thermodynamic profiles or short hairpin RNA (shRNA) vectors that generate double-stranded RNAs (dsRNAs), permitting processing specifically from either the 5' or 3' end of the incipient siRNA, results in the degradation of the respective target mRNAs of either strand of the siRNA duplex with comparable efficiencies. Thus, while most RNAi reactions may follow the thermodynamic asymmetry rule in strand selection, our study suggests an exceptional mode for certain siRNAs in which both strands of the duplex are competent in sponsoring RNAi, and implies additional factors that might dictate the RNAi targets.
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language English
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publisher Public Library of Science (PLoS)
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spelling doaj-art-743f2bfdc23241a79646ee06eda58d822025-08-20T02:33:43ZengPublic Library of Science (PLoS)PLoS ONE1932-62032009-01-0144e538210.1371/journal.pone.0005382Both strands of siRNA have potential to guide posttranscriptional gene silencing in mammalian cells.Jun-Xia WeiJie YangJi-Feng SunLin-Tao JiaYong ZhangHui-Zhong ZhangXia LiYan-Ling MengLi-Bo YaoAn-Gang YangDespite the widespread application of RNA interference (RNAi) as a research tool for diverse purposes, the key step of strand selection of siRNAs during the formation of RNA-induced silencing complex (RISC) remains poorly understood. Here, using siRNAs targeted to the complementary region of Survivin and the effector protease receptor 1 (EPR-1), we show that both strands of the siRNA duplex can find their target mRNA and are equally eligible for assembly into Argonaute 2 (Ago2) of RISC in HEK293 cells. Transfection of the synthetic siRNA duplexes with different thermodynamic profiles or short hairpin RNA (shRNA) vectors that generate double-stranded RNAs (dsRNAs), permitting processing specifically from either the 5' or 3' end of the incipient siRNA, results in the degradation of the respective target mRNAs of either strand of the siRNA duplex with comparable efficiencies. Thus, while most RNAi reactions may follow the thermodynamic asymmetry rule in strand selection, our study suggests an exceptional mode for certain siRNAs in which both strands of the duplex are competent in sponsoring RNAi, and implies additional factors that might dictate the RNAi targets.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0005382&type=printable
spellingShingle Jun-Xia Wei
Jie Yang
Ji-Feng Sun
Lin-Tao Jia
Yong Zhang
Hui-Zhong Zhang
Xia Li
Yan-Ling Meng
Li-Bo Yao
An-Gang Yang
Both strands of siRNA have potential to guide posttranscriptional gene silencing in mammalian cells.
PLoS ONE
title Both strands of siRNA have potential to guide posttranscriptional gene silencing in mammalian cells.
title_full Both strands of siRNA have potential to guide posttranscriptional gene silencing in mammalian cells.
title_fullStr Both strands of siRNA have potential to guide posttranscriptional gene silencing in mammalian cells.
title_full_unstemmed Both strands of siRNA have potential to guide posttranscriptional gene silencing in mammalian cells.
title_short Both strands of siRNA have potential to guide posttranscriptional gene silencing in mammalian cells.
title_sort both strands of sirna have potential to guide posttranscriptional gene silencing in mammalian cells
url https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0005382&type=printable
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