Small interfering RNAs containing full 2′-O-methylribonucleotide-modified sense strands display argonaute2/eIF2C2-dependent activity
Small interfering RNAs containing full 2′-O-methylribonucleotide-modified sense strands display argonaute2/eIF2C2-dependent activity
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DOI：
10.1261/rna.2150806
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发表时间：
2006-01-01
期刊：
影响因子：
4.5
通讯作者：
Baker, BF
中科院分区：
文献类型：
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作者：
Kraynack, BA;Baker, BF
RNA干扰（RNAi）是短干扰RNA（siRNA）引导互补单链RNA降解的过程。在这项研究中，我们研究了全链硫代磷酸酯（PS）骨架和2 '-O-甲基（2，-OMe）糖修饰对RNAi介导的沉默的影响。与以前的报道相反，我们已经鉴定了含有完整的2 '-OMe修饰的有义链的活性siRNA双链体，其显示出与类似序列的未修饰类似物相当的活性。这些修饰的siRNA的结构是其活性的主要决定因素，序列和骨架组成是次要的。我们进一步表明，通过使用生物素标记的siRNA和亲和标记的hAgo 2/eIF 2C 2，在有义链中含有完全2 '-OMe取代的siRNA双链体的活性由RNA诱导的沉默复合物（RISC）介导，并且可以通过选择性掺入这些修饰来调节链特异性负载（或结合）至hAgo 2。
RNA interference (RNAi) is a process by which short interfering RNAs (siRNAs) direct the degradation of complementary single-strand RNAs. In this study, we investigated the effects of full-strand phosphorothioate (PS) backbone and 2'-O-methyl (2,-OMe) sugar modifications on RNAi-mediated silencing. In contrast to previous reports, we have identified active siRNA duplexes containing full 2'-OMe-modified sense strands that display comparable activity to the unmodified analog of similar sequence. The structure of these modified siRNAs is the predominant determinant of their activity, with sequence and backbone composition being secondary. We further show, by using biotin-tagged siRNAs and affinity-tagged hAgo2/eIF2C2, that activity of siRNA duplexes containing full 2'-OMe substitutions in the sense strand is mediated by the RNA-induced silencing complex (RISC) and that strand-specific loading (or binding) to hAgo2 may be modulated through selective incorporation of these modifications.