Issue 2, 2021

Single-nucleotide resolution of N6-adenine methylation sites in DNA and RNA by nitrite sequencing

Abstract

A single-nucleotide resolution sequencing method of N6-adenine methylation sites in DNA and RNA is described. Using sodium nitrite under acidic conditions, chemoselective deamination of unmethylated adenines readily occurs, without competing deamination of N6-adenine sites. The deamination of adenines results in the formation of hypoxanthine bases, which are read by polymerases and reverse transcriptases as guanine; the methylated adenine sites resist deamination and are read as adenine. The approach, when coupled with high-throughput DNA sequencing and mutational analysis, enables the identification of N6-adenine sites in RNA and DNA within various sequence contexts.

Graphical abstract: Single-nucleotide resolution of N6-adenine methylation sites in DNA and RNA by nitrite sequencing

Supplementary files

Article information

Article type
Edge Article
Submitted
24 jun 2020
Accepted
04 nov 2020
First published
05 nov 2020
This article is Open Access

All publication charges for this article have been paid for by the Royal Society of Chemistry
Creative Commons BY license

Chem. Sci., 2021,12, 606-612

Single-nucleotide resolution of N6-adenine methylation sites in DNA and RNA by nitrite sequencing

Y. Mahdavi-Amiri, K. Chung Kim Chung and R. Hili, Chem. Sci., 2021, 12, 606 DOI: 10.1039/D0SC03509B

This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. You can use material from this article in other publications without requesting further permissions from the RSC, provided that the correct acknowledgement is given.

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