Chemical Approaches to Control the Function of Regulatory RNAs

控制调节性 RNA 功能的化学方法

• 批准号: 10581333
• 负责人: ERIKS ROZNERS
• 金额: $ 10.1万
• 依托单位: STATE UNIVERSITY OF NY,BINGHAMTON
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-02-01 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10581333
• 关键词: Address; Amides; Binding; Biological; Biological Models; Biological Sciences; Biology; Charge; Chemicals; Clustered Regularly Interspaced Short Palindromic Repeats; Collaborations; Complex; DNA; Development; Disease; Double-Stranded RNA; Electrostatics; Future; Goals; Hydrogen Bonding; Ligand Binding; Ligands; Medicine; MicroRNAs; Modification; Molecular Conformation; Oligonucleotides; Peptide Nucleic Acids; Pharmacologic Substance; Pharmacy (field); Positioning Attribute; Property; Proteins; RNA; RNA Binding; RNA Interference; Research; Small Interfering RNA; Specificity; Structure; Therapeutic; Untranslated RNA; Vertebral column; analog; design; genetic information; improved; inorganic phosphate; insight; novel; novel therapeutic intervention; novel therapeutics; programs; tool; triple helix; uptake

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 in biology and medicine. Our research program strives to capitalize on these exciting future discoveries by 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 projects involve collaborations with structural biochemists (Martin Egli), biological chemists (Naoki Sugimoto), and a pharmaceutical company (Alnylam). 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.

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，将继续提供生物学的新发现， 和医药我们的研究计划致力于利用这些令人兴奋的未来发现， 化学修饰以调节调节RNA的结构和功能。长期目标是 1）开发用于基础研究和生物医学应用的新型RNA化学修饰，以及 2)探索双链RNA（dsRNA）序列特异性识别的新模式。我们的研究 该计划包括两个不同但相互关联的项目：1）酰胺作为新的骨架修饰， 调节RNA，和2）修饰的肽核酸（PNA）对双链RNA的序列特异性识别。 项目1在短干扰RNA和RNA中用酰胺键取代核苷酸间磷酸 与成簇规则间隔短回文重复序列（CRISPR）相关。这些目标 改善这些RNA的细胞摄取、递送和序列特异性。前提是酰胺 可以模拟磷酸盐与蛋白质的结构和氢键相互作用，并且在某些位置， 能够重塑和改善这些互动。项目2探索化学修饰的PNA作为配体 用于生物医学上重要的dsRNA的序列特异性识别。我们的目标是改善细胞 摄取PNA，并证明使用microRNA作为起始肽的三链体形成的生物学效应。 模型系统前提是M-修饰的三链体形成PNA是唯一适合于序列- 这将使得能够识别生物学上重要的非编码dsRNA。 未来的研究将集中在CRISPR RNA的化学修饰和使用三螺旋来控制 复杂的非编码RNA的构象。这些项目涉及与结构生物化学家的合作 （马丁·埃格利），生物化学家（杉本直树）和一家制药公司（Alnylam）。两 项目有一个共同的主题，即设计利用电荷的化学修饰， RNA靶标与配体和与RNA相互作用的蛋白质之间的互补性。的 一个突破性的想法是开发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。