Multiplexed Analysis of the Epitranscriptome

项目摘要/摘要 目前已知有170多种天然发生的对RNA的化学修饰，其中100多种已被发现 在人类所有类型的RNA中：mRNA、tRNA、rRNA、lncRNA等。这些修改起着核心作用 在RNA功能的几乎所有方面，如翻译的启动和终止，翻译的保真度，选择性 剪接、细胞间的运输和调节RNA的降解。安装、阅读、 而去除修饰是目前制药行业最感兴趣的药物靶点。目前可用 分析方法要么不报告序列上下文，要么提供序列信息，但代价是 具有多路复用能力。尽管有这些限制，但现在已知修改动态地绑定到 癌症进展、耐药性和病毒感染的表型变化。此应用程序的重点是 为了降低一种新的RNA修饰分析方法的风险，该方法能够读取 多重反应，接近单碱基分辨率。这项技术将具有重要意义，因为它将 提供第一种方法，用于分析和关联整个 给定样本中的转录组。

Project Summary/Abstract More than 170 naturally occurring chemical modifications to RNA are known, more than 100 of which are found in human RNA of all types: mRNA, tRNA, rRNA, lncRNA, and the others. These modifications play a central role in nearly all aspects of RNA function, such as translation initiation and termination, translation fidelity, alternative splicing, trafficking between cellular compartments, and regulating RNA degradation. Proteins that install, read, and remove modifications are promising drug targets of high current interest to pharma. Currently available analytical methods either do not report on sequence context or provide sequence information but at the expense of multiplexing capability. Despite these limitations, it is now known that modifications are dynamically tied to phenotypic changes in cancer progression, drug resistance, and viral infection. The focus of this application is to de-risk a new approach to RNA modification analysis capable of reading any set of RNA modifications in a multiplexed reaction, approaching single base resolution. This technology will be significant because it will provide the first method for profiling and correlating changes of multiple RNA modification types across the entire transcriptome in a given sample.