Reconstitution of RNA Interference Machinery
Reconstitution of RNA Interference Machinery
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DOI：
10.1007/978-1-4939-7339-2_9
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发表时间：
2018-01-01
期刊：
影响因子：
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通讯作者：
Tomari, Yukihide
中科院分区：
文献类型：
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作者：
Iwasaki, Shintaro;Tomari, Yukihide
小RNA，包括小干扰RNAs(SiRNAs)和微小RNAs(MiRNAs)，通过形成称为RNA诱导沉默复合体(RISC)的效应复合体，从携带它们互补序列的靶mRNAs中沉默蛋白质表达。虽然RISC的组装机制已经研究了近20年，但具体的机制仍不清楚，部分原因是缺乏一个纯粹的重建系统。最近，我们鉴定了果蝇RISC组装所需的所有核心蛋白，并成功地用8种重组蛋白重述了具有催化活性的RISC组装。重建系统为RISC组装的详细研究提供了一个通用的框架，包括本期另一章中描述的单分子分析。
Small RNAs, including small interfering RNAs (siRNAs) and microRNAs (miRNAs), silence protein expression from target mRNAs bearing their complementary sequences, via the formation of the effector complex called RNA-induced silencing complex (RISC). Although the mechanism of RISC assembly has been studied for nearly two decades, the detailed mechanism has still remained unclear in part due to the lack of a pure reconstitution system. Recently, we identified all the core proteins necessary for RISC assembly in flies and successfully recapitulated the assembly of catalytically active RISC with eight recombinant proteins. The reconstitution system provides a versatile framework for detailed studies of RISC assembly, including single molecule analysis as described in another chapter in this issue.