Chekanova Julia A, Belostotsky Dmitry A
Department of Biological Sciences, State University of New York, Albany, NY, USA.
Methods Mol Biol. 2006;342:73-85. doi: 10.1385/1-59745-123-1:73.
Although initially believed to act exclusively as translational repressors, microRNAs (miRNAs) are now known to target complementary messenger RNA (mRNA) transcripts for either translational repression or cleavage via the RNA-induced silencing complex (RISC) ([1], reviewed in ref. 2). The current model postulates that mature miRNAs are incorporated into the RISC, bind target mRNAs based on complementarity, and guide cleavage of mRNA targets with perfect or nearly perfect complementarity and translational repression of targets with lower complementarity (2). The translational repression mechanism of miRNA-mediated gene regulation, which is common in animals but also exists in plants, is not well understood mechanistically. Conversely, miRNA-directed mRNA cleavage by RISC is common in plants, but also occurs in animals (3). This chapter focuses on the mRNA cleavage by miRNA-programmed RISC, and, specifically, on characterizing the products of such cleavage.
尽管最初认为微小RNA(miRNA)仅作为翻译抑制因子起作用,但现在已知它们通过RNA诱导沉默复合体(RISC)靶向互补信使RNA(mRNA)转录本,以实现翻译抑制或切割([1],参考文献2中有综述)。当前模型假定成熟的miRNA被整合到RISC中,基于互补性结合靶mRNA,并引导具有完美或近乎完美互补性的mRNA靶标切割以及具有较低互补性的靶标的翻译抑制(2)。miRNA介导的基因调控的翻译抑制机制在动物中很常见,但在植物中也存在,其机制尚未得到很好的理解。相反,RISC介导的miRNA指导的mRNA切割在植物中很常见,但在动物中也会发生(3)。本章重点关注miRNA编程的RISC对mRNA的切割,特别是对这种切割产物的表征。