Author:
Zhu Yinzhou,Pirnie Stephan P.,Carmichael Gordon G.
Abstract
Ribose methylation (2′-O-methylation, 2′-OMe) occurs at high frequencies in rRNAs and other small RNAs and is carried out using a shared mechanism across eukaryotes and archaea. As RNA modifications are important for ribosome maturation, and alterations in these modifications are associated with cellular defects and diseases, it is important to characterize the landscape of 2′-O-methylation. Here we report the development of a highly sensitive and accurate method for ribose methylation detection using next-generation sequencing. A key feature of this method is the generation of RNA fragments with random 3′-ends, followed by periodate oxidation of all molecules terminating in 2′,3′-OH groups. This allows only RNAs harboring 2′-OMe groups at their 3′-ends to be sequenced. Although currently requiring microgram amounts of starting material, this method is robust for the analysis of rRNAs even at low sequencing depth.
Funder
National Institute of General Medical Sciences
Publisher
Cold Spring Harbor Laboratory
Cited by
77 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献