Laboratoire Interdisciplinaire Carnot de Bourgogne, UMR 6303 CNRS, Université de Bourgogne, Dijon Cedex, France.
Laboratoire Léon Brillouin LLB, CEA, CNRS UMR12, Université Paris Saclay, Gif-Sur-Yvette, France.
Methods Mol Biol. 2020;2113:319-327. doi: 10.1007/978-1-0716-0278-2_20.
Atomic force and transmission electron microscopies (AFM/TEM) are powerful tools to analyze RNA-based nanostructures. While cryo-TEM analysis allows the determination of near-atomic resolution structures of large RNA complexes, this chapter intends to present how RNA nanostructures can be analyzed at room temperature on surfaces. Indeed, TEM and AFM analyses permit the conformation of a large population of individual molecular structures to be observed, providing a statistical basis for the variability of these nanostructures within the population. Nevertheless, if double-stranded DNA molecular imaging has been described extensively, only a few investigations of single-stranded DNA and RNA filaments have been conducted so far. Indeed, technique for spreading and adsorption of ss-molecules on AFM surfaces or TEM grids is a crucial step to avoid disturbing RNA conformation on the surface. In this chapter, we present a specific method to analyze RNA assemblies and RNA-protein complexes for molecular microscopies.
原子力显微镜和透射电子显微镜（AFM/TEM）是分析基于 RNA 的纳米结构的强大工具。虽然冷冻 TEM 分析可以确定大型 RNA 复合物的近原子分辨率结构，但本章旨在介绍如何在室温下在表面上分析 RNA 纳米结构。事实上，TEM 和 AFM 分析可以观察到大量单个分子结构的构象，为这些纳米结构在群体中的变异性提供了统计学基础。然而，如果双链 DNA 分子成像已经被广泛描述，那么到目前为止，只有少数关于单链 DNA 和 RNA 纤维的研究。事实上，在 AFM 表面或 TEM 网格上展开和吸附 ss 分子的技术是避免在表面上干扰 RNA 构象的关键步骤。在本章中，我们提出了一种用于分析 RNA 组装体和 RNA-蛋白质复合物的分子显微镜的特定方法。
