Chemical Approaches to Control the Function of Regulatory RNAs

近几十年来，我们对RNA的看法发生了巨大变化。虽然RNA最初被认为是 RNA是将遗传信息从DNA转移到蛋白质的被动信使，现在很清楚，RNA是 一个令人兴奋的和未充分探索的调节分子，将继续提供新的发现新的 生物学和医学的发现。我们的研究重点是利用化学修饰来调节 结构和功能调控RNA。长期目标是1）开发新型RNA化学物质 基础研究和生物医学应用的修改，以及2）探索新的序列模式- 特异性识别双链RNA（dsRNA）。我们的研究计划包括两个不同的，但 相关项目：1）酰胺作为调控RNA的新型骨架修饰，2）序列- 修饰的肽核酸（PNA）特异性识别dsRNA。项目1取代核苷酸间 在短干扰RNA和与成簇规则相关的RNA中具有酰胺键的磷酸盐 间隔短回文重复序列（interspaced short palindromic repeats，CRISPR）目标是改善细胞摄取、递送和 这些RNA的序列特异性。前提是酰胺可以模拟结构和氢键 磷酸盐与蛋白质的相互作用，并且在某些位置，可能能够重塑和改善 这些互动。项目2探索化学修饰的PNA作为序列特异性的配体， 识别生物医学上重要的dsRNA。目的是改善PNA的细胞摄取， 使用microRNA作为初始模型系统证明了三链体形成的生物学效应。的 前提是M-修饰的三链体形成PNA独特地适合于序列特异性识别 这将使得能够识别生物学上重要的非编码dsRNA。未来的研究将集中 CRISPR RNA的化学修饰和使用三螺旋控制复合物的构象 非编码RNA。这两个项目有一个共同的主题，即设计化学修饰， RNA靶标与配体和与其相互作用的蛋白质之间的电荷互补性的优点 核糖核酸这个超出范围的想法是开发RNA化学修饰和RNA结合配体， 无效的静电排斥和利用有效的静电吸引，同时 增强分子相互作用的序列特异性。这种推力源于我们最近的发现 RNA对中和磷酸盐负电荷的化学修饰反应异常敏感 在RNA本身和RNA结合寡核苷酸类似物中，如果成功，我们的研究将 有助于解决RNA干扰，CRISPR，识别治疗相关 RNA，并将为开发独特的研究工具和新的治疗策略打开大门。

Recent decades have dramatically changed our view of RNA. While RNA was initially believed to be barely a passive messenger in the transfer of genetic information from DNA to proteins, it is now clear that RNA is an exciting and underexplored regulatory molecule that will continue to deliver new discoveries new discoveries in biology and medicine. Our research is focused on using chemical modifications to modulate the structure and function regulatory RNAs. The long-term goals are to 1) develop novel RNA chemical modifications for fundamental studies and biomedical applications, and 2) explore new modes of sequence- specific recognition of double-stranded RNA (dsRNA). Our research program comprises two distinct but interrelated projects: 1) amides as novel backbone modifications for regulatory RNAs, and 2) sequence- specific recognition of dsRNA by modified peptide nucleic acids (PNA). Project 1 replaces internucleotide phosphates with amide linkages in short interfering RNAs and RNAs associated with clustered regularly interspaced short palindromic repeats (CRISPR). The goals are to improve the cellular uptake, delivery and sequence specificity of these RNAs. The premise is that amides can mimic structure and H-bonding interactions of phosphates with proteins and, at certain positions, may be able to remodel and improve these interactions. Project 2 explores chemically modified PNA as a ligand for sequence-specific recognition of biomedically important dsRNA. The goals are to improve the cellular uptake of PNA and to demonstrate the biological effect of triplex formation using microRNAs as the initial model system. The premise is that M-modified triplex-forming PNAs are uniquely suited for sequence-specific recognition of dsRNA and will enable recognition of biologically important non-coding dsRNA. Future research will focus on chemical modifications of CRISPR RNAs and using the triple helix to control conformations of complex non-coding RNAs. The two projects share a common theme of designing chemical modifications that take advantage of charge complementarity between the RNA target and the ligands and proteins interacting with RNA. The overreaching idea is to develop RNA chemical modifications and RNA binding ligands that avoid unproductive electrostatic repulsion and capitalize on productive electrostatic attraction while concurrently enhancing sequence specificity of molecular interactions. This thrust grows out of our recent discoveries that RNA is unusually receptive to chemical modifications that neutralize the negative charge of phosphate backbone, both in RNA itself and in RNA binding oligonucleotide analogues. If successful, our research will contribute to addressing key gaps in RNA interference, CRISPR, recognition of therapeutically relevant RNAs, and will open doors for development of unique research tools and new therapeutic strategies.