Architecture et Réactivité de l'ARN, Université de Strasbourg, CNRS, Strasbourg, France.
Wiley Interdiscip Rev RNA. 2019 Mar;10(2):e1518. doi: 10.1002/wrna.1518. Epub 2018 Nov 28.
RNA molecules are important players in all domains of life and the study of the relationship between their multiple flexible states and the associated biological roles has increased in recent years. For several decades, chemical and enzymatic structural probing experiments have been used to determine RNA structure. During this time, there has been a steady improvement in probing reagents and experimental methods, and today the structural biologist community has a large range of tools at its disposal to probe the secondary structure of RNAs in vitro and in cells. Early experiments used radioactive labeling and polyacrylamide gel electrophoresis as read-out methods. This was superseded by capillary electrophoresis, and more recently by next-generation sequencing. Today, powerful structural probing methods can characterize RNA structure on a genome-wide scale. In this review, we will provide an overview of RNA structural probing methodologies from a historical and technical perspective. This article is categorized under: RNA Structure and Dynamics > RNA Structure, Dynamics, and Chemistry RNA Methods > RNA Analyses in vitro and In Silico RNA Methods > RNA Analyses in Cells.
RNA 分子是所有生命领域的重要参与者，近年来，研究它们的多种灵活状态与相关生物学功能之间的关系变得越来越重要。几十年来，化学和酶学结构探测实验被用于确定 RNA 结构。在此期间，探测试剂和实验方法不断得到改进，如今，结构生物学家社区拥有广泛的工具，可以在体外和细胞中探测 RNA 的二级结构。早期的实验使用放射性标记和聚丙烯酰胺凝胶电泳作为读出方法。随后，毛细管电泳取代了它，最近又被下一代测序取代。如今，强大的结构探测方法可以在全基因组范围内对 RNA 结构进行特征分析。在这篇综述中，我们将从历史和技术的角度来概述 RNA 结构探测方法。本文属于以下分类：RNA 结构和动态>RNA 结构、动态和化学；RNA 方法>体外和计算机模拟的 RNA 分析；RNA 方法>细胞内的 RNA 分析。
