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.

项目概要/摘要 对RNA结构与功能关系的日益认识导致了对新RNA的需求 能够靶向特定 RNA 结构的技术。这些技术对于 开发用于研究 RNA 功能的探针和治疗 RNA 介导的疾病的疗法。然而， 除了与核糖体结合的抗生素之外，结构特异性 RNA 结合试剂非常罕见。因此， 开发能够实现 RNA 结构特异性靶向的新技术仍然是一个重要的挑战 在许多领域。 我的研究项目的中心愿景是解决结构特异性 RNA 结合的缺陷 试剂使用完全不同类型的核酸亲和试剂：L-适体。 L-适体是独一无二的 因为它们由基于 L-（脱氧）核糖的核酸（L-DNA 和 L-RNA）组成，它们是镜像 天然 D-核苷酸的图像（对映体）。因为寡核苷酸具有相反的立体化学（D 与 L) 无法形成连续的 Watson-Crick 碱基对，我们能够进化 L-适体通过三级相互作用（形状）而不是自适应地结合结构化 D-RNA 靶标 初级序列。换句话说，L-适体摆脱了 Watson-Crick 碱基配对的束缚，从而实现了 更细致的分子识别模式有待发现。结果，L-适体结合结构化RNA 与传统的亲和试剂相比，具有更高的亲和力和特异性。结合 RNA 基于其 形状而不是沃森克里克碱基配对代表了与传统的显着背离 基于寡核苷酸的方法代表了适体技术的重大进步。 未来五年，我的研究小组计划进一步开发L-适体技术，以便 实现其作为实用研究和治疗工具的承诺。特别是，我们将重点关注纳入 修饰的核苷酸赋予 L-适体类似蛋白质的功能，从而产生一类新型的 RNA 靶向抗体模拟物。因为这些技术的发展都是在这样的背景下进行的 疾病相关 RNA，例如致癌 microRNA 和病毒 RNA，这项工作将立即产生影响 通过产生先导试剂来探索疾病的病因并开发新的治疗策略来产生影响。在 与我的愿景一致，我们的目标是确定 L-适体-D-RNA 复合物的结构，这将提供 深入了解这种新颖的识别模式并为未来的 L-适体设计提供信息。