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因子。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驱动的，这些技术能够准确检测、映射和操作这些标记。在这里，我们总结了核酸修饰研究的最新技术进展，重点介绍了能够准确检测和映射这些修饰的技术。对于讨论的每个修饰（N ⁶ -甲基腺苷、5-甲基胞苷、肌苷、假尿苷和N ⁴-乙酰胞苷），我们首先介绍其映射和检测的“金标准”技术，然后讨论为解决金标准的任何缺点而开发的技术。通过强调这些技术的共性和不同之处，我们希望提供对该领域当前状态的看法，并为未来技术的发展制定指导方针。