Nano-Antibodies for Studying RNA Modifications

用于研究 RNA 修饰的纳米抗体

• 批准号: 8841036
• 负责人: Jiwu Wang
• 金额: $ 22.5万
• 依托单位: ALLELE BIOTECHNOLOGY AND PHARMACEUTICALS
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-30 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8841036
• 关键词: 2' -O-methyluridine; 8-methylguanosine; Affinity; Alleles; Animals; Antibodies; Antigens; Area; Belgium; Benchmarking; Binding; Biological Assay; Carrier Proteins; Coin; Communities; Conjugated Carrier; Development; Disease; Dyes; Escherichia coli; Family; Foundations; Functional RNA; Gene Expression Regulation; Goals; Grant; Haptens; Image; Immune; Immunoprecipitation; Legal patent; Llama; Marketing; Measures; Messenger RNA; Methylation; Modification; Molecular and Cellular Biology; Monitor; Monoclonal Antibodies; National Institute of Drug Abuse; National Institute of Mental Health; Nature; Netherlands; Nucleosides; Nurses; Oryctolagus cuniculus; Outcome; Paper; Pharmacologic Substance; Phase; Property; Proteins; Psyche structure; Publishing; RNA; Reagent; Research; Research Personnel; Research Project Grants; Resolution; Ribose; Shark; Small Business Innovation Research Grant; Specificity; Substance abuse problem; System; Technology; Test Result; Testing; United States National Institutes of Health; Variant; base; drug candidate; experience; expiration; improved; innovation; nano; novel; phase 2 study; polyclonal antibody; prototype; public health relevance; success; tool

DESCRIPTION (provided by applicant): The goal of the proposed Phase I research is to develop a set of commercial reagents for recognizing 2'-O-methyladenosine (Am), 2'-O-methylcytidine (Cm), 2'-O-methylguanosine (Gm) or 2'-O-methyluridine (Um), as well as indiscriminately to all four 2'-O-methyl ribose moiety. The core technologies that serve as the foundation of these new reagents are based on the unique properties of camelid antibodies, which have been well accepted as valuable binding reagents for proteins and haptens, but so far without any example for RNA modifications. The main advantage of the proposed single-domain (VHH) antibodies from camelidae is that they have very high affinity and specificity, and they are easy to reproduce as monoclonal antibodies, stable, and of very small size, enabling super-resolution imaging when conjugated to dye. Currently, affinity reagents of any type are only available for monitoring a handful among the more than 100 RNA modification moieties. The best examples of such available reagents are the antibodies against N6-methyladenosine (m6A), which proved to be of significant value to researchers in the field trying to understand the functions of the m6A-modified mRNAs. However, the two anti-m6A antibodies, created about 3 decades ago, are both rabbit polyclonal antibodies that are not easily reproducible or suitable for high precision immune-imaging or other modern assays. There is an urgent need for development of high quality antibodies against the fast-expanding and increasingly important family of known modified mRNAs and regulatory RNAs. Successful execution of the proposed project will establish a fully tested, market-ready group of reagents and kits for detecting one type of closely related RNA modifications, 2'-O-methylation on the ribose of the 4 RNA bases. Currently, there is a lack of such reagents on the market, providing the applicant a very good chance of commercial success with the outcome of the proposed project. After achieving this goal, similar efforts can be initiated in a phase II SBIR or other types of project to develop more nano-antibodies against additional RNA modifications, eventually overcome the bottleneck of post-transcriptional RNA studies, and serve as the foundation for advancing NIDA- and NIMH-relevant research projects on RNA influence in various substance abuse or mental diseases.

DESCRIPTION (provided by applicant): The goal of the proposed Phase I research is to develop a set of commercial reagents for recognizing 2'-O-methyladenosine (Am), 2'-O-methylcytidine (Cm), 2'-O-methylguanosine (Gm) or 2'-O-methyluridine (Um), as well as indiscriminately to all four 2'-O-methyl ribose moiety.作为这些新试剂基础的核心技术是基于骆驼抗体的独特特性，这些特性已被很好地接受为蛋白质和触觉的有价值的结合试剂，但到目前为止，没有任何RNA修饰的例子。 Camelidae提出的单域（VHH）抗体的主要优点是它们具有很高的亲和力和特异性，并且它们易于繁殖，因为单克隆抗体，稳定且尺寸很小，可以在连接到染料时实现超分辨率成像。当前，任何类型的亲和力试剂仅可用于监测100多个RNA修饰部分中的少数。这种可用试剂的最佳例子是针对N6-甲基腺苷（M6A）的抗体，事实证明，这对于试图了解该领域的研究人员具有重要价值 M6A修饰mRNA的功能。但是，大约3年前创建的两种抗M6A抗体都是兔多克隆抗体，它们不容易再现或适合于高精度免疫成像或其他现代测定。迫切需要开发针对快速扩张和越来越重要的已知改性mRNA和监管RNA的家族的高质量抗体。拟议项目的成功执行将建立一个经过全面测试的，可以进行市场准备就绪的试剂和套件，用于检测一种紧密相关的RNA修饰，在4个RNA碱基的核糖上进行2'-O-甲基化。目前，市场上缺乏此类试剂，这为申请人提供了拟议项目的结果很有可能获得商业成功的机会。实现了这一目标之后，可以在II阶段SBIR或其他类型的项目中发起类似的努力，以开发更多 针对其他RNA修饰的纳米抗体，最终克服了转录后RNA研究的瓶颈，并成为推进NIDA和NIMH相关的研究项目的基础，该项目对RNA在各种药物滥用或精神疾病中的影响。