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.

描述(申请人提供)：拟议的第一阶段研究的目标是开发一套商业试剂，用于识别2‘-O-甲基腺苷(AM)、2’-O-甲基胞苷(Cm)、2‘-O-甲基鸟苷(GM)或2’-O-甲基尿苷(Um)，以及不分青红皂白地识别所有四个2‘-O-甲基核糖部分。作为这些新试剂基础的核心技术是基于骆驼抗体的独特性质，骆驼抗体已被广泛认为是有价值的蛋白质和半抗原结合试剂，但到目前为止还没有任何用于RNA修饰的例子。所提出的驼科单域(VHH)抗体的主要优点是它们具有非常高的亲和力和特异性，并且它们很容易作为单抗复制，稳定，并且尺寸非常小，当与染料结合时能够进行超分辨率成像。目前，任何类型的亲和试剂都只能用于监测100多个RNA修饰部分中的一小部分。这类试剂最好的例子是针对N6-甲基腺苷(M6A)的抗体，事实证明，这种抗体对该领域的研究人员试图了解 M6A修饰的mRNA的功能。然而，这两种抗m6A抗体产生于大约30年前，都是兔多克隆抗体，不易复制，也不适合高精度免疫成像或其他现代分析。迫切需要开发针对快速扩张和日益重要的已知修饰RNA和调节RNA家族的高质量抗体。拟议项目的成功执行将建立一组经过充分测试、市场准备就绪的试剂和试剂盒，用于检测一种密切相关的RNA修饰，即4个RNA碱基核糖上的2‘-O-甲基化。目前，市场上缺乏这样的试剂，这为申请人提供了一个非常好的机会，以拟议的项目的结果在商业上成功。在实现这一目标后，可以在第二阶段SBIR或其他类型的项目中启动类似的努力，以开发更多 针对额外RNA修饰的纳米抗体，最终克服转录后RNA研究的瓶颈，并作为推进NIDA和NIMH相关研究项目的基础，研究RNA在各种药物滥用或精神疾病中的影响。