Targeted mRNA degradation by double-stranded RNA in vitro
Targeted mRNA degradation by double-stranded RNA in vitro
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DOI：
10.1101/gad.13.24.3191
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发表时间：
1999-12-15
影响因子：
10.5
通讯作者：
Sharp, PA
中科院分区：
文献类型：
--
作者：
Tuschl, T;Zamore, PD;Sharp, PA
双链RNA（dsRNA）在包括脊椎动物在内的许多生物中指导基因特异性的转录后沉默，并且为研究基因功能提供了新的工具。这种dsRNA干扰（RNAi）的生化机制尚不清楚。在这里，我们报告了一个无细胞系统的发展，从合胞胚盘果蝇胚胎，概括了许多RNAi的功能。该反应中观察到的干扰是序列特异性的，由双链RNA而不是单链RNA促进，通过特定的mRNA降解发挥作用，并且需要最小长度的双链RNA。此外，dsRNA的预孵育增强了其活性。这些结果表明，RNAi可以通过可溶性反应中的序列特异性过程介导。
Double-stranded RNA (dsRNA) directs gene-specific, post-transcriptional silencing in many organisms, including vertebrates, and has provided a new tool for studying gene function. The biochemical mechanisms underlying this dsRNA interference (RNAi) are unknown. Here we report the development of a cell-free system from syncytial blastoderm Drosophila embryos that recapitulates many of the features of RNAi. The interference observed in this reaction is sequence specific, is promoted by dsRNA but not single-stranded RNA, functions by specific mRNA degradation, and requires a minimum length of dsRNA. Furthermore, preincubation of dsRNA potentiates its activity. These results demonstrate that RNAi can be mediated by sequence-specific processes in soluble reactions.