Probing RNA structure and dynamics using cutting-edge biophysical techniques

RNA plays a central role in numerous cellular processes, from gene expression to catalysis and regulation. Understanding the structural dynamics of RNA is crucial for elucidating its diverse functional mechanisms. In this review, we highlight the latest advancements in biophysical techniques that have provided unprecedented insights into the complex conformational landscapes and time-resolved dynamics of RNA molecules. We begin by discussing the application of nuclear magnetic resonance (NMR) spectroscopy, which has emerged as a powerful tool for probing RNA structure and dynamics at atomic resolution. We describe how NMR-based approaches, including relaxation dispersion and chemical exchange saturation transfer (CEST) experiments, have revealed the existence of sparsely populated, transient RNA conformations that are essential for biological function. Next, we explore the use of single-molecule techniques, such as optical tweezers and nanopore sequencing, to study RNA dynamics in real-time and under physiologically relevant conditions. These methods have shed light on the kinetics and mechanisms of RNA folding, as well as the role of RNA-binding proteins and small molecules in modulating RNA structure and dynamics. Finally, we discuss the integration of computational approaches, including molecular dynamics simulations and machine learning algorithms, with experimental data to develop a more comprehensive understanding of RNA dynamics across multiple timescales. By combining cutting-edge biophysical tools with advanced computational methods, researchers are poised to uncover the intricate relationship between RNA structure, dynamics, and function. Overall, this review highlights the remarkable progress made in probing the structural dynamics of RNA, paving the way for new discoveries and a deeper understanding of this versatile biomolecule.