RNAs play different functional roles in human biology and disease dysfunction.For the vast majority of RNA molecules,a complex high-level structure is a prerequisite for their function.Understanding the three-dimensional(3D)structures of non-coding RNAs is essential for studying their functions,pathogenesis,and developing targeted drugs.However,due to their inherent flexibilities,few RNA 3D structures have been determined experimentally.Cryo-electron microscopy(cryo-EM)has stood out among other structural analysis methods in recent years thanks to the rapid development of detector technology and software algorithms and is not constrained by the requirements of NMR for small molecular weights and X-ray crystallography for crystallization.In this review,we summarize the recent progress of RNA 3D structure analysis by cryo-EM in combination with other techniques(e.g.,nuclear magnetic resonance,crystallography,and theoretical modeling).The objective of this review is to facilitate a comprehensive understanding of RNA structures,enabling improved investigations into this intricate molecular composition and the development of RNA-targeted therapeutic interventions.
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