Gu Sam G, Pak Julia, Barberan-Soler Sergio, Ali Mustapha, Fire Andrew, Zahler Alan M
Department of Molecular Cell and Developmental Biology, Center for Molecular Biology of RNA, University of California Santa Cruz 95064, USA.
RNA. 2007 Sep;13(9):1492-504. doi: 10.1261/rna.581907. Epub 2007 Jul 24.
MicroRNAs (miRNAs) are regulatory molecules that share both biosynthetic derivation (cleavage from short hairpin precursor RNAs) and functional roles (downregulation of specific mRNAs through targeted degradation and/or translational inhibition). A distinct family of small RNAs, termed siRNAs, have some common characteristics but exhibit distinct modes of biosynthesis and function. In this study, we report procedures for purification of a predominant species of miRNA-containing ribonucleoprotein complexes from Caenorhabditis elegans and demonstrate that this population is distinct from the predominant pool of siRNA-containing ribonucleoprotein complexes. An observed miRNP-associated RNA population consisting predominantly (>95%) of miRNAs supported the unique identity of miRNPs as biological effectors within the cell, provided clean material for analysis of changes in miRNA spectra during development, and provided strong evidence of miRNA character for a number of novel small RNAs. Likewise, the RNA spectrum derived from partial siRNP purification was useful in defining functional characteristics of this more diverse population of small RNAs.
微小RNA（miRNA）是一类调控分子，它们在生物合成来源（从短发夹前体RNA切割而来）和功能作用（通过靶向降解和/或翻译抑制下调特定mRNA）方面具有共性。另一类被称为小干扰RNA（siRNA）的独特小RNA家族具有一些共同特征，但在生物合成和功能模式上表现出明显差异。在本研究中，我们报告了从秀丽隐杆线虫中纯化一种主要的含miRNA核糖核蛋白复合物的方法，并证明该群体与主要的含siRNA核糖核蛋白复合物群体不同。观察到的主要由miRNA组成（>95%）的与miRNP相关的RNA群体支持了miRNP作为细胞内生物效应物的独特身份，为分析发育过程中miRNA谱的变化提供了纯净材料，并为许多新型小RNA的miRNA特性提供了有力证据。同样，从部分siRNP纯化得到的RNA谱有助于定义这类更多样化的小RNA群体的功能特征。
