Recent advances in detection techniques for RNA modifications

<p indent="0mm">RNA is a single-stranded macromolecule that consists of adenosine (A), guanosine (G), uridine (U), and cytidine (C) nucleotides. It has been demonstrated that RNA can participate in the flow of genetic information and regulate gene expression. RNA modifications, referring to the addition of chemical groups at specific positions, can increase the complexity and modify the function of RNA molecules. These modifications are widely found in various types of RNA, including messenger RNA (mRNA), long non-coding RNA (lncRNA), transfer RNA (tRNA), ribosomal RNA (rRNA), and micro RNA (miRNA) in both prokaryotes and eukaryotes. Previous studies have shown that RNA undergoes extensive dynamic and reversible modifications that participate in biological processes, including gene expression regulation, RNA translation, cell differentiation, and disease development. Over 160 different types of RNA modifications have been identified through biological research, including N¹-methyladenosine (m¹A), N⁴-acetylcytidine (ac⁴C), N6-methyladenosine (m⁶A), N⁷-methylguanosine (m⁷G), 2ʹ-<italic>O</italic>-dimethyladenosine (m⁶Am), 2ʹ-<italic>O</italic>-methylated nucleosides (Nm), 5-methylcytosine (m⁵C), 5-hydroxymethylcytosine (hm⁵C), inosine (I), and pseudouridine (Ψ). Additionally, research on the functions of RNA modifications has been promoted by the development of detection techniques. Therefore, techniques that detect RNA modifications can provide evidence for a comprehensive understanding of the epigenetics and offer powerful tools for functional studies. Over the past several decades, researchers have developed unique methods to detect the various forms and distribution of RNA modifications. Existing detection techniques for RNA modifications can be grouped into three categories based on different detection principles, including quantification methods, site-specific detection methods, and sequencing methods. Due to the diversity of chemical modifications in RNA metabolism and functional regulation, detection techniques for RNA modification provide comprehensive insights into the modification status of the transcriptome and further lay the foundation for studying the functions of RNA modifications. In this review, we systematically introduce the principles, procedures, benefits, and drawbacks of different detection techniques for RNA modifications. Moreover, we summarize the latest research progress in RNA modification detection techniques, providing better insights and approaches to further study the biological functions and mechanisms of RNA modifications. Taken together, the continuous growth of detection techniques will improve the study of the functions and mechanisms of RNA modifications, providing a theoretical basis for further exploring the roles of RNA modifications in epigenetics. Therefore, it is crucial to develop more efficient, high-throughput, and high-resolution detection methods for detection techniques of RNA modifications in the future.