The detection, function, and therapeutic potential of RNA 2'-O-methylation

RNA modifications play crucial roles in shaping RNA structure, function, and metabolism. Their dysregulation has been associated with many diseases, including cancer, developmental disorders, cardiovascular diseases, as well as neurological and immune-related conditions. A particular type of RNA modification, 2'-O-methylation (Nm) stands out due to its widespread occurrence on all four types of nucleotides (A, U, G, C) and in most RNA categories, e.g., mRNA, rRNA, tRNA, miRNA, snRNA, snoRNA, and viral RNA. Nm is the addition of a methyl group to the 2' hydroxyl of the ribose moiety of a nucleoside. Given its great biological significance and reported association with many diseases, we first reviewed the occurrences and functional implications of Nm in various RNA species. We then summarized the reported Nm detection methods, ranging from biochemical techniques in the 70’s and 80’s to recent methods based on Illumina RNA sequencing, artificial intelligence (AI) models for computational prediction, and the latest nanopore sequencing methods currently under active development. Moreover, we discussed the applications of Nm in the realm of RNA medicine, highlighting its therapeutic potential. At last, we present perspectives on potential research directions, aiming to offer insights for future investigations on Nm modification. highlight the previously underappreciated roles for Nm in regulating RNA molecules beyond the traditional understanding of its function in non-coding RNAs. The enzymes responsible for the addition of the methyl group are known as RNA methyltransferases. More than a dozen proteins, including methyltransferases and their co-factors, have been reported to play roles in the installation or modulation of Nm in diverse RNA molecules (Table 1). Despite the wide involvement of these proteins in complex diseases such as cancer, Alzheimer’s, autoimmune diseases, and many others,³ the exact role of Nm in these pathological conditions remains to be explored. Understanding the mechanisms and implications of Nm will provide valuable insights into the molecular basis of these diseases and may contribute to the development of novel therapeutic strategies. Although less extensively studied compared to some other modifications such as the m⁶A and m⁵C, significant research progress has been made in Nm in the past decade. This significant progress benefits largely from advancements in detection technology based on the next-generation sequencing (NGS) methods and nanopore sequencing technologies. In this review, we first introduce the occurrences and functions of Nm in different RNA categories. We then provide a comprehensive overview of the methods employed to detect Nm in RNAs. Finally, we introduce the applications and discuss the future perspectives of Nm investigations.