Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Mahdavi-Amiri, Yasaman, Chung Kim Chung, Kimberley, Hili, Ryan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Royal Society of Chemistry 2020
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
_version_ 1783703692346654720
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
work_keys_str_mv AT mahdaviamiriyasaman singlenucleotideresolutionofn6adeninemethylationsitesindnaandrnabynitritesequencing
AT chungkimchungkimberley singlenucleotideresolutionofn6adeninemethylationsitesindnaandrnabynitritesequencing
AT hiliryan singlenucleotideresolutionofn6adeninemethylationsitesindnaandrnabynitritesequencing