Discovery and Function of Higher-Order RNA Structure

摘要 RNA在所有细胞中都是信息交换的中心通道。Rna分子对此进行编码。 在它们的初级序列和当RNA折叠回来时形成的复杂结构中的信息 单打独斗。到目前为止，我们由Mira支持的工作有一个始终如一的主题：生物功能调节 By RNA结构在真细菌和真核生物中普遍存在。博大精深 RNA结构的根本重要性是显而易见的，但仍然存在着深刻的知识鸿沟：我们 一般只对大部分地区的RNA结构有基本的了解 信使和非编码RNA。我们需要超越对RNA结构的建模，直接 检测细胞中的穿透空间结构通信和相互作用。我们的实验室采用了近乎独一无二的 方法，同时注重实验的简单性和直接性以及广泛性 模型系统中的验证。在基本上每一种情况下，我们都应用了严格和 研究RNA结构-功能相互关系的定量技术，关于 生物调控机制应运而生。我们还表明，具有更高- 订单结构更有可能包含高信息含量的缝隙和口袋，这些缝隙和口袋将小 分子，为以RNA为靶标的药物发现提供了一个充满活力的领域。这项工作的目的是 通过追求三个最重要的机会，产生了持续的长期影响。首先，项目重点是 开发决定性的、简明的和广泛可实现的技术来发现和 评估RNA中的高阶结构。其次，该项目直接和实验地应用了这些 从识别关键结构到广泛重要的挑战的简明方法 复制致病病毒以了解RNA构象集合是如何支配的 转录基因调控和前信使核糖核酸剪接。第三，理解RNA的易感性 形成更高层次的结构--通常是裂缝和口袋--打开了以这些结构为目标的可能性 具有小分子的主题，我们将利用这个机会，长期目标是发明易用和 创建RNA靶向疗法的简单策略。

SUMMARY RNA functions as the central conduit of information exchange in all cells. RNA molecules encode this information in both their primary sequences and in complex structures that form when RNA folds back on itself. A consistent theme from our MIRA-supported work to date is that biological function mediated by RNA structure is ubiquitous in both eubacterial and eukaryotic organisms. The broad and fundamental importance of RNA structure is clear, but there remains a profound knowledge gap: We generally have only rudimentary understanding of RNA structure across vast regions of most messenger and non-coding RNAs. We need to move beyond modeling RNA structure to directly detecting through-space structural communication and interactions in cells. Our lab takes a near-unique approach, simultaneously focusing on experimental simplicity and directness and on extensive validation in model systems. In essentially every instance where we have applied rigorous and quantitative technologies to study RNA structure-function interrelationships, new insights regarding biological regulatory mechanisms have emerged. We have also shown that RNA elements with higher- order structures are more likely to contain high-information-content clefts and pockets that bind small molecules, broadly informing a vigorous field of RNA-targeted drug discovery. This work is designed to have sustained long-term impact by pursuing three overarching opportunities. First, the project focuses on development of decisive, concise, and broadly implementable technologies for discovering and assessing higher-order structure in RNA. Second, this project applies these direct and experimentally concise methods to challenges of broad importance, from identification of structures essential for replication of pathogenic viruses to understanding how RNA conformational ensembles govern transcriptional gene regulation and pre-mRNA splicing. Third, understanding the propensity of RNA to form higher-order structures – and often clefts and pockets – opens up the possibility of targeting these motifs with small molecules, an opportunity we will exploit with the long-term goal of inventing facile and straightforward strategies for creating RNA-targeted therapeutics.