Department of Molecular Biosciences, University of Texas at Austin, 2500 Speedway, 78712 Austin TX, USA.
Brief Funct Genomics. 2021 Mar 27;20(2):77-85. doi: 10.1093/bfgp/elaa024.
Nearly 200 distinct chemical modifications of RNAs have been discovered to date. Their analysis via direct methods has been possible in abundant RNA species, such as ribosomal, transfer or viral RNA, since several decades. However, their analysis in less abundant RNAs species, especially cellular messenger RNAs, was rendered possible only recently with the advent of high throughput sequencing techniques. Given the growing biomedical interest of the proteins that write, erase and read RNA modifications, ingenious new methods to enrich and identify RNA modifications at base resolution have been implemented, and more efforts are underway to render them more quantitative. Here, we review several crucial modification-specific (bio)chemical approaches and discuss their advantages and shortcomings for exploring the epitranscriptome.
迄今为止，已经发现了近 200 种不同的 RNA 化学修饰。几十年来，通过直接方法已经可以对核糖体 RNA、转移 RNA 或病毒 RNA 等丰富的 RNA 进行分析。然而，直到最近高通量测序技术的出现，才使得对丰度较低的 RNA 物种，特别是细胞信使 RNA 的分析成为可能。鉴于 RNA 修饰的写入、擦除和读取蛋白的日益增长的生物医学兴趣，已经开发出了巧妙的新方法来以碱基分辨率富集和鉴定 RNA 修饰，并且正在进行更多的努力以提高它们的定量水平。在这里，我们综述了几种关键的修饰特异性（生物）化学方法，并讨论了它们在探索转录组学中的优缺点。
