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Single-nucleotide resolution of N(6)-adenine methylation sites in DNA and RNA by nitrite sequencing
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 ad...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Royal Society of Chemistry
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8179008/ https://www.ncbi.nlm.nih.gov/pubmed/34163791 http://dx.doi.org/10.1039/d0sc03509b |
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author | Mahdavi-Amiri, Yasaman Chung Kim Chung, Kimberley Hili, Ryan |
author_facet | Mahdavi-Amiri, Yasaman Chung Kim Chung, Kimberley Hili, Ryan |
author_sort | Mahdavi-Amiri, Yasaman |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-8179008 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | The Royal Society of Chemistry |
record_format | MEDLINE/PubMed |
spelling | pubmed-81790082021-06-22 Single-nucleotide resolution of N(6)-adenine methylation sites in DNA and RNA by nitrite sequencing Mahdavi-Amiri, Yasaman Chung Kim Chung, Kimberley Hili, Ryan Chem Sci Chemistry 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. The Royal Society of Chemistry 2020-11-05 /pmc/articles/PMC8179008/ /pubmed/34163791 http://dx.doi.org/10.1039/d0sc03509b Text en This journal is © The Royal Society of Chemistry https://creativecommons.org/licenses/by/3.0/ |
spellingShingle | Chemistry Mahdavi-Amiri, Yasaman Chung Kim Chung, Kimberley Hili, Ryan Single-nucleotide resolution of N(6)-adenine methylation sites in DNA and RNA by nitrite sequencing |
title | Single-nucleotide resolution of N(6)-adenine methylation sites in DNA and RNA by nitrite sequencing |
title_full | Single-nucleotide resolution of N(6)-adenine methylation sites in DNA and RNA by nitrite sequencing |
title_fullStr | Single-nucleotide resolution of N(6)-adenine methylation sites in DNA and RNA by nitrite sequencing |
title_full_unstemmed | Single-nucleotide resolution of N(6)-adenine methylation sites in DNA and RNA by nitrite sequencing |
title_short | Single-nucleotide resolution of N(6)-adenine methylation sites in DNA and RNA by nitrite sequencing |
title_sort | single-nucleotide resolution of n(6)-adenine methylation sites in dna and rna by nitrite sequencing |
topic | Chemistry |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8179008/ https://www.ncbi.nlm.nih.gov/pubmed/34163791 http://dx.doi.org/10.1039/d0sc03509b |
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