检测线粒体转录组中的 5-醛基胞嘧啶

Detection of 5-formylcytosine in Mitochondrial Transcriptome.

机构信息

Medical Research Council Mitochondrial Biology Unit, University of Cambridge, Cambridge, UK.

出版信息

Methods Mol Biol. 2021;2192:59-68. doi: 10.1007/978-1-0716-0834-0_5.

Abstract

Posttranscriptional RNA modifications have recently emerged as essential posttranscriptional regulators of gene expression. Here we present two methods for single nucleotide resolution detection of 5-formylcytosine (fC) in RNA. The first relies on chemical protection of fC against bisulfite treatment, the second method is based on chemical reduction of fC to hmC. In combination with regular bisulfite treatment of RNA, the methods allow for precise mapping of fC. The protocol is used for fC detection in mtDNA-encoded RNA, however, it can be straightforwardly applied for transcriptome-wide analyses.

摘要

近年来，转录后 RNA 修饰已成为基因表达的重要转录后调控因子。本文介绍了两种用于单核苷酸分辨率检测 RNA 中 5-甲酰胞嘧啶 (fC) 的方法。第一种方法依赖于化学保护 fC 免受亚硫酸氢盐处理，第二种方法基于 fC 化学还原为羟甲基胞嘧啶 (hmC)。与 RNA 的常规亚硫酸氢盐处理相结合，这些方法允许对 fC 进行精确定位。该方案用于检测 mtDNA 编码 RNA 中的 fC，但可以直接应用于全转录组分析。