Université de Strasbourg, CNRS, Architecture et Réactivité de l'ARN, UPR9002, F-67000 Strasbourg, France.
Viruses. 2021 Sep 22;13(10):1894. doi: 10.3390/v13101894.
RNA molecules are key players in a variety of biological events, and this is particularly true for viral RNAs. To better understand the replication of those pathogens and try to block them, special attention has been paid to the structure of their RNAs. Methods to probe RNA structures have been developed since the 1960s; even if they have evolved over the years, they are still in use today and provide useful information on the folding of RNA molecules, including viral RNAs. The aim of this review is to offer a historical perspective on the structural probing methods used to decipher RNA structures before the development of the selective 2'-hydroxyl acylation analyzed by primer extension (SHAPE) methodology and to show how they have influenced the current probing techniques. Actually, these technological breakthroughs, which involved advanced detection methods, were made possible thanks to the development of next-generation sequencing (NGS) but also to the previous works accumulated in the field of structural RNA biology. Finally, we will also discuss how high-throughput SHAPE (hSHAPE) paved the way for the development of sophisticated RNA structural techniques.
RNA 分子是各种生物事件中的关键参与者，病毒 RNA 更是如此。为了更好地了解这些病原体的复制并试图阻止它们，人们特别关注它们的 RNA 结构。自 20 世纪 60 年代以来，已经开发出了探测 RNA 结构的方法；即使这些方法多年来一直在不断发展，它们今天仍在使用，并为 RNA 分子（包括病毒 RNA）的折叠提供了有用的信息。本综述的目的是提供在选择性 2'-羟基酰化分析引物延伸 (SHAPE) 方法发展之前用于破译 RNA 结构的结构探测方法的历史视角，并展示它们如何影响当前的探测技术。实际上，这些涉及先进检测方法的技术突破之所以成为可能，要归功于下一代测序 (NGS) 的发展，也要归功于结构 RNA 生物学领域的前期工作积累。最后，我们还将讨论高通量 SHAPE（hSHAPE）如何为复杂的 RNA 结构技术的发展铺平道路。
