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.

中文翻译：

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病毒 RNA 的化学和酶学探测：从婴儿期到成熟期及以后

RNA 分子在各种生物事件中发挥着关键作用，对于病毒 RNA 来说尤其如此。为了更好地了解这些病原体的复制并尝试阻止它们，人们特别关注它们的 RNA 结构。自 20 世纪 60 年代以来，人们就开发出了探测 RNA 结构的方法；即使它们经过了多年的进化，今天仍然在使用，并提供有关 RNA 分子（包括病毒 RNA）折叠的有用信息。本综述的目的是在开发引物延伸分析选择性 2'-羟基酰化 (SHAPE) 方法之前，对用于破译 RNA 结构的结构探测方法提供历史视角，并展示它们如何影响当前的探测技术。事实上，这些涉及先进检测方法的技术突破之所以成为可能，不仅得益于下一代测序（NGS）的发展，也得益于之前在结构RNA生物学领域积累的工作成果。最后，我们还将讨论高通量 SHAPE (hSHAPE) 如何为复杂 RNA 结构技术的发展铺平道路。