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Single-nucleotide resolution of N ⁶-adenine methylation sites in DNA and RNA by nitrite sequencing.

ABSTRACT: A single-nucleotide resolution sequencing method of N ⁶-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 N ⁶-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 N ⁶-adenine sites in RNA and DNA within various sequence contexts.

SUBMITTER: Mahdavi-Amiri Y

PROVIDER: S-EPMC8179008 | biostudies-literature | 2020 Nov

REPOSITORIES: biostudies-literature

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Single-nucleotide resolution of N 6-adenine methylation sites in DNA and RNA by nitrite sequencing.

Mahdavi-Amiri Yasaman Y Chung Kim Chung Kimberley K Hili Ryan R

Chemical science 20201105 2

A single-nucleotide resolution sequencing method of N 6-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 N 6-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 deamin ...[more]

PMID: 34163791

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