Department of Chemistry and Biochemistry, University of North Carolina Greensboro, Greensboro, NC, USA.
Joint School of Nanoscience and Nanoengineering, University of North Carolina Greensboro, Greensboro, NC, USA.
Methods Mol Biol. 2024;2822:335-349. doi: 10.1007/978-1-0716-3918-4_21.
Every chemical group that is added to any one of the canonical ribonucleotides in a transcript would create a specific RNA modification. Currently, 170+ RNA modifications have been identified. A specific epitranscriptome refers to all the RNA modifications in a given biological system and is considered to play an important role in the regulations of cellular activities. Mass spectrometry-based methods have proven to be the most accurate way to identify RNA modifications and determine the amount of each detectable modification. Relating to the recent development of mapping specific RNA modifications within a transcriptome, the profiling of all RNA modifications can serve as a prescreening tool for mapping and provides support for analyzing the data obtained from mapping. In this chapter, the details for setting up a commonly used mass spectrometry-based method to profile all the RNA modifications in specific epitranscriptomes are described, and the possible options if available are discussed.
每一种添加到转录本中任何一个规范核苷酸的化学基团都会产生一种特定的 RNA 修饰。目前已经鉴定出 170 多种 RNA 修饰。特定的 RNA 修饰组是指给定生物系统中的所有 RNA 修饰，被认为在细胞活动的调控中发挥重要作用。基于质谱的方法已被证明是鉴定 RNA 修饰和确定每种可检测修饰物的最准确方法。与最近在转录组中定位特定 RNA 修饰的发展有关，对所有 RNA 修饰的分析可以作为定位的预筛选工具，并为分析从定位获得的数据提供支持。本章描述了建立一种常用的基于质谱的方法来分析特定的 RNA 修饰组中所有 RNA 修饰的详细信息，并讨论了如果有可用选项的话。
