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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Public Library of Science (PLoS)
2009
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oai:doaj.org-article:743f2bfdc23241a79646ee06eda58d822021-11-25T06:22:56ZBoth strands of siRNA have potential to guide posttranscriptional gene silencing in mammalian cells.1932-620310.1371/journal.pone.0005382https://doaj.org/article/743f2bfdc23241a79646ee06eda58d822009-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/19401777/?tool=EBIhttps://doaj.org/toc/1932-6203Despite 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.Jun-Xia WeiJie YangJi-Feng SunLin-Tao JiaYong ZhangHui-Zhong ZhangXia LiYan-Ling MengLi-Bo YaoAn-Gang YangPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 4, Iss 4, p e5382 (2009) |
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Medicine R Science Q 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. |
| 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. |
| format |
article |
| 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 |
| title |
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_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_sort |
both strands of sirna have potential to guide posttranscriptional gene silencing in mammalian cells. |
| publisher |
Public Library of Science (PLoS) |
| publishDate |
2009 |
| url |
https://doaj.org/article/743f2bfdc23241a79646ee06eda58d82 |
| work_keys_str_mv |
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| _version_ |
1718413807602106368 |