The 2-O-methylation of the ribose moiety is one of the most common RNA modifications and is present in diverse highly abundant non-coding RNAs (ncRNAs) and at the 5 cap of mRNAs. Moreover, many studies have revealed that Nm at 3’-end is presented in some ncRNAs, such as microRNAs (miRNAs) in plants as well as PIWI-interacting RNAs (piRNAs) in animals.This modification can perturb the function of ribosomes and disrupt tRNA decoding, regulate alternative splicing fidelity, protect ncRNAs from 3’-5’ exonucleolytic degradation and provide a molecular signature for discrimination of self from non-self mRNA.