It has become increasingly evident that the structures RNAs adopt are conformationally dynamic; the various structured states that RNAs sample govern their interactions with other nucleic acids, proteins, and ligands to regulate a myriad of biological processes. Although several biophysical approaches have been developed and used to study the dynamic landscape of structured RNAs, technical limitations have limited their application to all classes of RNA due to variable size and flexibility. Recent advances combining chemical probing experiments with next-generation- and direct sequencing have emerged as an alternative approach to exploring the conformational dynamics of RNA. In this review, we provide a methodological overview of the sequencing-based techniques used to study RNA conformational dynamics. We discuss how different techniques have enabled us to better understand the propensity of RNAs from a variety of different classes to sample multiple conformational states. Finally, we present examples of the ways these techniques have reshaped how we think about RNA structure.

中文翻译：

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使用纳米孔和下一代测序探测 RNA 结构和动力学


越来越明显的是，RNA 采用的结构是动态构象的。 RNA 采样的各种结构状态控制着它们与其他核酸、蛋白质和配体的相互作用，从而调节无数的生物过程。尽管已经开发了几种生物物理方法并用于研究结构化 RNA 的动态景观，但由于大小和灵活性的可变性，技术限制限制了它们对所有类别 RNA 的应用。化学探测实验与下一代直接测序相结合的最新进展已成为探索 RNA 构象动力学的替代方法。在这篇综述中，我们提供了用于研究 RNA 构象动力学的基于测序的技术的方法学概述。我们讨论了不同的技术如何使我们能够更好地了解各种不同类别的 RNA 对多种构象状态进行采样的倾向。最后，我们举例说明这些技术如何重塑我们对 RNA 结构的看法。