RNA甲基化修饰检测研究进展

Advances in the detection of RNA epigenetic modifications

摘要:

RNA修饰是转录后调控基因表达的重要因素，在表观遗传学领域具有重要的研究意义。目前，许多已知的RNA修饰检测方法依赖抗体的特异性。最常用的是基于RNA甲基化免疫沉淀结合测序法（methylated RNA immunoprecipitation sequencing，MeRIP-seq），但这种方法也容易产生假阳性和假阴性结果。因此，近年来出现了一系列免抗体的测序方法，以及探索成本低廉、简单易操作的免抗体RNA修饰生物传感检测新方法。此外，特定位点的RNA修饰检测是探明甲基化对疾病具体调控机制的关键，单碱基分辨检测方法构建是必要的前提。本综述总结近年来RNA甲基化修饰特异性检测以及特定位点RNA修饰定量检测的新方法。主要从基于免疫途径、甲基化特异性酶途径、杂交方法、化学处理、标记技术以及RNA修饰原位检测技术进行总结归纳，分析不同检测方法的优点和不足。同时讨论了通用性检测方法及第三代测序用于直接检测RNA甲基化修饰的研究前景。

Abstract:

RNA modification is an important factor in the post-transcriptional regulation of gene expression and is of great research importance in all areas of epigenetics. Currently，many known methods for RNA modification detection rely in part on the specificity of antibodies. The most commonly used method is high-throughput sequencing based on antibody immunoprecipitation，but this method is also prone to false-positive and false-negative results. Therefore，a series of antibody-free sequencing methods have also emerged in recent years，as well as the exploration of new methods for cost-effective and simple antibody-free RNA modification biosensing assays. In addition，site-specific RNA modification detection is crucial for probing the specific regulatory mechanisms of methylation on diseases，and single-base resolution assay construction is a necessary prerequisite. This review summarizes new methods for the specific detection of RNA methylation modifications in recent years and for the quantitative detection of site-specific RNA modifications，and mainly summarize the advantages and shortcomings of different detection methods based on immune pathways，methylation-specific enzyme pathways，hybridization methods，chemical treatment，labeling techniques，and in situ detection of RNA modifications. We also discuss the research prospects of universal detection methods and third-generation sequencing for direct detection of RNA methylation modifications.