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学科主题: 药学
题名:
Focusing on RISC assembly in mammalian cells
作者: 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
发表日期: 2008-04-11
DOI: 10.1016/j.bbrc.2008.01.116
卷: 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
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/68211
Appears in Collections:北京大学药学院_期刊论文

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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

Recommended Citation:
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.
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