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Focusing on RISC assembly in mammalian cells
Hong, Junmei1; Wei, Na1; Chalk, Alistair2; Wang, Jue1; Song, Yutong3; Yi, Fan1; Qiao, Ren-Ping4; Sonnharnmer, Erik L. L.5; Wahlestedt, Claes6; Liang, Zicai1,2; Du, Quan1
关键词siRNA RISC assembly silencing activity thermodynamic stability
刊名BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
2008-04-11
DOI10.1016/j.bbrc.2008.01.116
368期:3页:703-708
收录类别SCI
文章类型Article
WOS标题词Science & Technology
类目[WOS]Biochemistry & Molecular Biology ; Biophysics
研究领域[WOS]Biochemistry & Molecular Biology ; Biophysics
关键词[WOS]SMALL INTERFERING RNAS ; SIRNA SEQUENCES ; ASYMMETRY ; EFFICACY ; DATABASE ; DUPLEXES ; COMPLEX ; DESIGN
英文摘要

RISC (RNA-induced silencing complex) is a central protein complex in RNAi, into which a siRNA strand is assembled to become effective in gene silencing. By using an in vitro RNAi reaction based on Drosophila embryo extract, an asymmetric model was recently proposed for RISC assembly of siRNA strands, suggesting that the strand that is more loosely paired at its 5 ′ end is selectively assembled into RISC and results in target gene silencing. However, in the present study, we were unable to establish such a correlation in cell-based RNAi assays, as well as in large-scale RNAi data analyses. This suggests that the thermodynamic stability of siRNA is not a major determinant of gene silencing in mammalian cells. Further studies on fork siRNAs showed that mismatch at the 5 ′ end of the siRNA sense strand decreased RISC assembly of the antisense strand, but surprisingly did not increase RISC assembly of the sense strand. More interestingly, measurements of melting temperature showed that the terminal stability of fork siRNAs correlated with the positions of the mismatches, but not gene silencing efficacy. In summary, our data demonstrate that there is no definite correlation between siRNA stability and gene silencing in mammalian cells, which suggests that instead of thermodynamic stability, other features of the siRNA duplex contribute to RISC assembly in RNAi. (c) 2008 Elsevier Inc. All rights reserved.

语种英语
WOS记录号WOS:000253925900040
引用统计
被引频次:7[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.bjmu.edu.cn/handle/400002259/68211
专题北京大学药学院
作者单位1.Peking Univ, Inst Mol Med, Beijing 100871, Peoples R China
2.Karolinska Inst, Dept Mol Med & Surg, S-17176 Stockholm, Sweden
3.Karolinska Inst, Dept Woman & Child Hlth, S-17176 Stockholm, Sweden
4.Peking Univ, Sch Pharmaceut Sci, State Key Lab Nat & Biomimet Drugs, Beijing 100083, Peoples R China
5.Stockholm Bioinformat Ctr, S-17174 Stockholm, Sweden
6.Scripps Florida, Jupiter, FL 33458 USA
推荐引用方式
GB/T 7714
Hong, Junmei,Wei, Na,Chalk, Alistair,et al. Focusing on RISC assembly in mammalian cells[J]. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS,2008,368(3):703-708.
APA Hong, Junmei.,Wei, Na.,Chalk, Alistair.,Wang, Jue.,Song, Yutong.,...&Du, Quan.(2008).Focusing on RISC assembly in mammalian cells.BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS,368(3),703-708.
MLA Hong, Junmei,et al."Focusing on RISC assembly in mammalian cells".BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 368.3(2008):703-708.
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