RNA三维结构以及结构-功能关系是目前的学术前沿和热点课题。RNA柔性大、难结晶，其结构研究严重滞后于其他生物大分子，亟需发展新的结构分析手段。此项目中，我们将以建立高效、快速、普适的RNA三维结构解析的氢检测固体核磁共振方法学为研究目标。以HIV病毒二聚引发位点RNA为模型，发展适用于固体核磁共振研究的非结晶RNA固体样品的制备方法，提高固体核磁共振研究RNA的普适性；发展基于PAIN和PAR机制的，长程距离约束采集实验技术，提高结构分析的研究效率；以及建立快速魔角旋转条件下、三维氢检测固体核磁共振实验方法。通过设计的三维固体核磁共振实验，并结合同位素标记策略和快速魔角旋转实验技术，建立RNA结构的氢检测固体核磁共振方法学体系，并完成HIV病毒二聚引发位点RNA片段的结构解析。本课题深入发展固体核磁共振研究RNA结构的实验技术，为未知RNA三维结构解析提供新的研究思路。

Structure determination on large RNAs is a formidable challenge in the field of structural biology. Most RNAs contain flexible segments and are difficult to form large 3D crystals, limiting the applications of traditional structural biology tools. The development of RNA structures largely lag behind that of other biomacromolecues. This project aims in the methodologies advancement of RNA structure characterization by solid-state NMR spectroscopy. A RNA fragment corresponding to the dimerization initiation site of the HIV genome will be applied as a model system to develop solid-state NMR methods. We will develop the non-crystalline RNA solid-state sample preparation approach. Selectively 2D labeling approaches joint with fast magic angle spinning solid-state NMR spectroscopy will be applied to develop the three dimensional proton-detected solid-state NMR spectroscopy and to facilitate in collecting long-range distance restraints for structure determination. This project will advance multiple solid-state NMR methods, which would support RNA structure characterization.