Mirror Image Aptamers: Next Generation RNA-Binding Reagents for Basic Research and Therapeutic Applications

镜像适体：用于基础研究和治疗应用的下一代 RNA 结合试剂

• 批准号: 9382491
• 负责人: Jonathan Thomas Sczepanski
• 金额: $ 36.24万
• 依托单位: TEXAS A&M UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-15 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9382491
• 关键词: Address; Affinity; Antibiotics; Antibodies; Aptamer Technology; Base Pairing; Basic Science; Binding; Cellular biology; Complex; DNA; Development; Disease; Drug Targeting; Etiology; Future; Image; Lead; Mediating; MicroRNAs; Nucleic Acids; Nucleotides; Oligonucleotides; Oncogenic; Play; Proteins; RNA; RNA Binding; Reagent; Research; Ribose; Ribosomes; Role; Shapes; Specificity; Structure; Structure-Activity Relationship; Technology; Therapeutic; Vision; Work; aptamer; base; design; drug development; enantiomer; innovation; insight; mimetics; molecular recognition; new technology; next generation; novel; novel therapeutic intervention; programs; stereochemistry; targeted treatment; three dimensional structure; tool; viral RNA

Project Summary/Abstract The increasing appreciation of RNA's structure-function relationship has led to a demand for new technologies that enable targeting of specific RNA structures. Such technologies are essential for the development of probes to study RNA function and therapeutics to treat RNA-mediated diseases. However, outside of antibiotics binding the ribosome, structure-specific RNA-binding reagents are very rare. Thus, developing of new technologies that enable structure-specific targeting of RNA remains an important challenge in many fields. The central vision of my research program is to address the deficit of structure-specific RNA-binding reagents using a radically different type of nucleic acid affinity reagent: L-aptamers. L-Aptamers are unique because they are comprised of L-(deoxy)ribose-based nucleic acids (L-DNA and L-RNA), which are mirror images (enantiomers) of natural D-nucleotides. Because oligonucleotides of opposite stereochemistry (D versus L) are incapable of forming contiguous Watson-Crick base pairs with each other, we are able to evolve L-aptamers that adaptively bind structured D-RNA targets through tertiary interactions (shape) rather than primary sequence. In other words, L-aptamers escape the tyranny of Watson-Crick base pairing, enabling a more nuanced mode of molecular recognition to be discovered. As a result, L-aptamers bind structured RNAs with greater affinity and specificity compared to conventional affinity reagent. Binding RNAs based on their shape rather than Watson-Crick base pairing represents a significant departure from traditional oligonucleotide-based approaches and represents a major advance in aptamer technology. During the next five year, my research group aims to further develop L-aptamer technology in order to realize its promise as a practical research and therapeutic tool. In particular, we will focus on incorporation of modified nucleotides that bestow protein-like functionality on L-aptamers, thus generating a novel class of RNA-targeted antibody mimetics. Because these technological developments will be carried out in the context of disease associated RNAs, such as oncogenic microRNAs and viral RNAs, this work will have an immediate impact by generating lead reagents to probe the etiology of disease and develop new therapeutic strategies. In line with my vision, we aim to determine the structure of an L-aptamer–D-RNA complex, which will provide insight into this novel mode of recognition and inform future L-aptamer design.

项目摘要/摘要 对RNA结构功能关系的越来越多的欣赏导致了对新的需求 能够靶向特定RNA结构的技术。这样的技术对于 研究RNA功能和治疗以治疗RNA介导的疾病的问题的发展。然而， 在结合核糖体的抗生素之外，结构特异性的RNA结合试剂非常罕见。那， 开发实现特定于结构的RNA的新技术仍然是一个重要的挑战 在许多领域。 我的研究计划的中心愿景是解决结构特定的RNA结合的确定 使用根本不同类型的核酸亲和力试剂的试剂：l-Aptamers。 L-Aptamers是独一无二的 因为它们是l-（脱氧）核糖基核酸（L-DNA和L-RNA）的积累，它们是镜子 天然D-核苷酸的图像（对映异构体）。因为相反立体化学的寡核苷酸（D） 与l）无法彼此形成连续的Watson-Crick Base对，我们能够进化 通过第三纪相互作用（形状）而不是 主要序列。换句话说，L-Appamers逃脱了Watson-Crick Base配对的暴政，使A 要发现的更细微的分子识别模式。结果，l-Appamer结合结构的RNA 与常规亲和力试剂相比，具有更大的亲和力和特异性。根据他们的绑定RNA 形状而不是沃森 - 克里克底座配对代表了与传统的显着差异 基于寡核苷酸的方法，代表了ATAMER技术的重大进步。 在接下来的五年中，我的研究小组旨在进一步开发L-Appamer技术，以便 实现其作为实践研究和治疗工具的承诺。特别是，我们将专注于 修饰的核动肽在L-Aptamers上赋予蛋白质样功能，从而产生一类新的 靶向RNA的抗体模拟物。因为这些技术发展将在上下文中进行 与疾病相关的RNA，例如致癌microRNA和病毒RNA，这项工作将立即具有 通过产生铅试剂来探究疾病的病因并制定新的治疗策略来影响。 与我的视野一致，我们旨在确定L-Appamer-D-RNA复合物的结构，该复合物将提供 深入了解这种新颖的识别方式，并为未来的L-Aptamer设计提供信息。