Single-stranded antisense siRNAs guide target RNA cleavage in RNAi

被引：1134

作者：

Martinez, J ^{[1
]}

Patkaniowska, A ^{[1
]}

Urlaub, H ^{[1
]}

Lührmann, R ^{[1
]}

Tuschl, T ^{[1
]}

机构：

[1] Max Planck Inst Biophys Chem, Dept Cellular Biochem, D-37077 Gottingen, Germany

来源：

CELL | 2002年 / 110卷 / 05期

关键词：

D O I：

10.1016/S0092-8674(02)00908-X

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Small interfering RNAs (siRNAs) are the mediators of mRNA degradation in the process of RNA interference (RNAi). Here, we describe a human biochemical system that recapitulates siRNA-mediated target RNA degradation. By using affinity-tagged siRNAs, we demonstrate that a single-stranded siRNA resides in the RNA-induced silencing complex (RISC) together with eIF2C1 and/or eIF2C2 (human GERp95) Argonaute proteins. RISC is rapidly formed in HeLa cell cytoplasmic extract supplemented with 21 nt siRNA duplexes, but also by adding single-stranded antisense RNAs, which range in size between 19 and 29 nucleotides. Single-stranded antisense siRNAs are also effectively silencing genes in HeLa cells, especially when 5'-phosphorylated, and expand the repertoire of RNA reagents suitable for gene targeting.

引用

页码：563 / 574

页数：12

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