Enzyme-mediated chemical modifications of nucleic acids are indispensable regulators of gene expression. Our understanding of the biochemistry and biological significance of these modifications has largely been driven by an ever-evolving landscape of technologies that enable accurate detection, mapping, and manipulation of these marks. Here we provide a summary of recent technical advances in the study of nucleic acid modifications with a focus on techniques that allow accurate detection and mapping of these modifications. For each modification discussed (N⁶-methyladenosine, 5-methylcytidine, inosine, pseudouridine, and N⁴-acetylcytidine), we begin by introducing the “gold standard” technique for its mapping and detection, followed by a discussion of techniques developed to address any shortcomings of the gold standard. By highlighting the commonalities and differences of these techniques, we hope to provide a perspective on the current state of the field and to lay out a guideline for development of future technologies.

酶介导的核酸化学修饰是基因表达不可或缺的调节剂。我们对这些修饰的生物化学和生物学意义的理解很大程度上是由不断发展的技术推动的，这些技术能够准确检测、绘制和操纵这些标记。在这里，我们总结了核酸修饰研究的最新技术进展，重点是能够准确检测和绘制这些修饰的技术。对于所讨论的每种修饰（ N ^{6 -}甲基腺苷、5-甲基胞苷、肌苷、假尿苷和N ^{4 -}乙酰胞苷），我们首先介绍其绘图和检测的“黄金标准”技术，然后讨论为解决这些问题而开发的技术金本位制的任何缺点。通过强调这些技术的共性和差异，我们希望为该领域的现状提供一个视角，并为未来技术的发展制定指南。