Department of Chemistry, University of North Carolina, Chapel Hill, NC 27599-3290, USA.
Department of Biology, University of North Carolina, Chapel Hill, NC 27599, USA.
Curr Opin Struct Biol. 2024 Oct;88:102877. doi: 10.1016/j.sbi.2024.102877. Epub 2024 Jul 17.
RNA molecules fold to form complex internal structures. Many of these RNA structures populate ensembles with rheostat-like properties, with each state having a distinct function. Until recently, analysis of RNA structures, especially within cells, was limited to modeling either a single averaged structure or computationally-modeled ensembles. These approaches obscure the intrinsic heterogeneity of many structured RNAs. Single-molecule correlated chemical probing (smCCP) strategies are now making it possible to measure and deconvolute RNA structure ensembles based on efficiently executed chemical probing experiments. Here, we provide an overview of fundamental single-molecule probing principles, review current ensemble deconvolution strategies, and discuss recent applications to diverse biological systems. smCCP is enabling a revolution in understanding how the plasticity of RNA structure is exploited in biological systems to respond to stimuli and alter gene function. The energetics of RNA ensembles are often subtle and a subset can likely be targeted to modulate disease-associated biological processes.
RNA 分子折叠形成复杂的内部结构。这些 RNA 结构中的许多结构都具有变阻器样的特性，每个状态都具有独特的功能。直到最近，RNA 结构的分析，特别是在细胞内，仅限于对单个平均结构或计算建模的结构进行分析。这些方法掩盖了许多结构 RNA 的固有异质性。现在，基于高效执行的化学探测实验，单分子相关化学探测（smCCP）策略正使得测量和解析 RNA 结构集合成为可能。在这里，我们提供了对基本单分子探测原理的概述，回顾了当前的集合反卷积策略，并讨论了最近在各种生物系统中的应用。smCCP 正在推动人们对 RNA 结构可塑性如何在生物系统中被利用来响应刺激和改变基因功能的理解发生革命性的变化。RNA 集合的能量通常很微妙，可能可以靶向一小部分来调节与疾病相关的生物过程。
