Next-generation computational/chemical methods for complex RNA structures

项目摘要 非编码RNA的不断发现及其在细胞和生物学中的关键作用 病毒机制正在激发新的抗菌，抗肿瘤，抗病毒和基因组编辑 基于禁用，操纵和重新利用相关RNA的疗法。 不幸的是，我们对“RNA如何工作”的生物物理学理解不足，阻碍了对RNA的研究。 这些潜在的拯救生命的努力。一个关键的瓶颈是， 晶体学、核磁共振、低温电子显微镜、系统发育分析 和生物化学方法来确定非编码的部分有序构象 所有功能状态的RNA。为了解决这个瓶颈，我们正在开发 实验方法和互补的计算方法， 足以推断和工程化RNA二级和三级结构的信息， 他们的异质性集合，通过社区范围内的盲法试验进行评估， 前瞻性补偿突变/拯救实验和全局RNA设计 挑战在这里，我们概述了我们的研究，将严格解决四个扩展 生物医学上的重大问题，迄今为止，在分子生物学上的进展有限， 建模工作：RNA结构在其天然细胞和病毒中的异质性 RNA与蛋白质和其他生物相互作用的建模和设计 分子，通过化学修饰进行调节; RNA的高精度计算 折叠能量学;以及动态3D RNA结构的自动化设计， 医疗应用。我们将通过持续的盲法试验来评估成功与否， 由十多位生物和生物工程专家进行的独立测试 合作者，并通过采用我们的方法和软件工具，由更广泛的 研究社区。就像专门的结构生物学工具和 计算机设计正在建立对蛋白质行为的坚定理解， 监管，我们建议，这里概述的技术将改变我们的 了解非编码RNA的结构，为他们的研究提供更强的基础。 生物医学激活或破坏。

PROJECT SUMMARY The continuing discoveries of non-coding RNAs and their critical roles in cellular and viral machinery are inspiring novel antibacterial, antitumor, antiviral, and genome-editing therapies based on disabling, manipulating, and repurposing the RNAs involved. Unfortunately, our poor biophysical understanding of `how RNAs work' hinders the development of these potentially life-saving efforts. A critical bottleneck has been the inapplicability of crystallography, NMR, cryoelectron microscopy, phylogenetic analysis, and biochemical methods to determine the partly ordered conformations of non-coding RNAs in all their functional states. To resolve this bottleneck, we are developing experimental methods and complementary computational approaches that give rich information sufficient to infer and engineer RNA secondary and tertiary structures and their heterogeneous ensembles, evaluated through community-wide blind trials, prospective compensatory mutation/rescue experiments, and global RNA design challenges. Here, we outline expansions of our research that will rigorously address four biomedically significant problems that have so far seen limited progress in molecular modeling efforts: the heterogeneity of RNA structures within their native cellular and viral contexts; modeling and design of RNA's biological interactions with proteins and other molecules, modulated by chemical modification; high-accuracy calculation of RNA folding energetics; and the automated design of dynamic 3D RNA structures for eventual medical applications. We will evaluate success through continuing blind trials, independent tests by more than a dozen expert biological and bioengineering collaborators, and through adoption of our methods and software tools by the broader research community. In the same way that specialized structural biology tools and computational design are establishing a firm understanding of protein behavior and regulation, we propose that the technologies outlined here will transform our understanding of structure in non-coding RNAs, providing a stronger basis for their biomedical activation or disruption.