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: 10001546
• 负责人: Jonathan Thomas Sczepanski
• 金额: $ 35.59万
• 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/10001546
• 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-功能关系的日益认识导致了对新的 能够以特定RNA结构为靶点的技术。这些技术对于 研究核糖核酸功能和治疗核糖核酸介导性疾病的探针的发展。然而， 除了抗生素与核糖体结合外，结构特异的RNA结合试剂非常少见。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。 我的研究计划的中心愿景是解决结构特定的RNA结合的缺陷 使用一种完全不同类型的核酸亲和试剂的试剂：L适体。L-适配子是独一无二的 因为它们是由L脱氧核糖基核酸(L脱氧核糖核酸和L核糖核酸)组成的，它们是镜像 天然D-核苷酸的图像(对映体)。因为具有相反立体化学结构的寡核苷酸(D VS L)不能彼此形成连续的沃森-克里克碱基对，我们能够进化 L-适体通过三级相互作用(形状)而不是 主序列。换句话说，L-适配子逃脱了沃森-克里克碱基配对的专制，使 更细微的分子识别模式有待发现。结果，L-适配子结合了结构化RNA 与传统的亲和试剂相比，具有更高的亲和力和特异性。基于其属性的绑定RNA 形状而不是沃森-克里克碱基配对代表着与传统 以寡核苷酸为基础的方法代表了适体技术的重大进步。 在接下来的五年里，我的研究小组的目标是进一步开发L适体技术，以便 实现其作为实用研究和治疗工具的前景。特别是，我们将专注于成立 修饰的核苷酸赋予L-适配子类似蛋白质的功能，从而产生一类新的 核糖核酸靶向抗体模拟。因为这些技术发展将在 对于疾病相关的RNA，如致癌的microRNA和病毒RNA，这项工作将立即产生 通过产生铅试剂来探索疾病的病因和开发新的治疗策略的影响。在……里面 与我的愿景一致，我们的目标是确定L-适体-D-RNA复合体的结构，这将为 洞察这种新颖的识别模式，并为未来的L-适配子设计提供信息。