Fast and precise detection of DNA methylation with tetramethylammonium-filled nanopore

Abstract The tremendous demand for detecting methylated DNA has stimulated intensive studies on developing fast single-molecule techniques with excellent sensitivity, reliability, and selectivity. However, most of these methods cannot directly detect DNA methylation at single-molecule level, which n...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Ying Wang, Yani Zhang, Yanli Guo, Xiao-feng Kang
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/d9155eca816146ab9618e01d64b28e49
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:d9155eca816146ab9618e01d64b28e49
record_format	dspace
spelling	oai:doaj.org-article:d9155eca816146ab9618e01d64b28e492021-12-02T12:32:56ZFast and precise detection of DNA methylation with tetramethylammonium-filled nanopore10.1038/s41598-017-00317-22045-2322https://doaj.org/article/d9155eca816146ab9618e01d64b28e492017-03-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-00317-2https://doaj.org/toc/2045-2322Abstract The tremendous demand for detecting methylated DNA has stimulated intensive studies on developing fast single-molecule techniques with excellent sensitivity, reliability, and selectivity. However, most of these methods cannot directly detect DNA methylation at single-molecule level, which need either special recognizing elements or chemical modification of DNA. Here, we report a tetramethylammonium-based nanopore (termed TMA-NP) sensor that can quickly and accurately detect locus-specific DNA methylation, without bisulfite conversion, chemical modification or enzyme amplification. In the TMA-NP sensor, TMA-Cl is utilized as a nanopore-filling electrolyte to record the ion current change in a single nanopore triggered by methylated DNA translocation through the pore. Because of its methyl-philic nature, TMA can insert into the methylcytosine-guanine (mC-G) bond and then effectively unfasten and reduce the mC-G strength by 2.24 times. Simultaneously, TMA can increase the stability of A-T to the same level as C-G. The abilities of TMA (removing the base pair composition dependence of DNA strands, yet highly sensing for methylated base sites) endow the TMA-NP sensor with high selectivity and high precision. Using nanopore to detect dsDNA stability, the methylated and unmethylated bases are easily distinguished. This simple single-molecule technique should be applicable to the rapid analysis in epigenetic research.Ying WangYani ZhangYanli GuoXiao-feng KangNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-10 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Ying Wang Yani Zhang Yanli Guo Xiao-feng Kang Fast and precise detection of DNA methylation with tetramethylammonium-filled nanopore
description	Abstract The tremendous demand for detecting methylated DNA has stimulated intensive studies on developing fast single-molecule techniques with excellent sensitivity, reliability, and selectivity. However, most of these methods cannot directly detect DNA methylation at single-molecule level, which need either special recognizing elements or chemical modification of DNA. Here, we report a tetramethylammonium-based nanopore (termed TMA-NP) sensor that can quickly and accurately detect locus-specific DNA methylation, without bisulfite conversion, chemical modification or enzyme amplification. In the TMA-NP sensor, TMA-Cl is utilized as a nanopore-filling electrolyte to record the ion current change in a single nanopore triggered by methylated DNA translocation through the pore. Because of its methyl-philic nature, TMA can insert into the methylcytosine-guanine (mC-G) bond and then effectively unfasten and reduce the mC-G strength by 2.24 times. Simultaneously, TMA can increase the stability of A-T to the same level as C-G. The abilities of TMA (removing the base pair composition dependence of DNA strands, yet highly sensing for methylated base sites) endow the TMA-NP sensor with high selectivity and high precision. Using nanopore to detect dsDNA stability, the methylated and unmethylated bases are easily distinguished. This simple single-molecule technique should be applicable to the rapid analysis in epigenetic research.
format	article
author	Ying Wang Yani Zhang Yanli Guo Xiao-feng Kang
author_facet	Ying Wang Yani Zhang Yanli Guo Xiao-feng Kang
author_sort	Ying Wang
title	Fast and precise detection of DNA methylation with tetramethylammonium-filled nanopore
title_short	Fast and precise detection of DNA methylation with tetramethylammonium-filled nanopore
title_full	Fast and precise detection of DNA methylation with tetramethylammonium-filled nanopore
title_fullStr	Fast and precise detection of DNA methylation with tetramethylammonium-filled nanopore
title_full_unstemmed	Fast and precise detection of DNA methylation with tetramethylammonium-filled nanopore
title_sort	fast and precise detection of dna methylation with tetramethylammonium-filled nanopore
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/d9155eca816146ab9618e01d64b28e49
work_keys_str_mv	AT yingwang fastandprecisedetectionofdnamethylationwithtetramethylammoniumfillednanopore AT yanizhang fastandprecisedetectionofdnamethylationwithtetramethylammoniumfillednanopore AT yanliguo fastandprecisedetectionofdnamethylationwithtetramethylammoniumfillednanopore AT xiaofengkang fastandprecisedetectionofdnamethylationwithtetramethylammoniumfillednanopore
_version_	1718393949333225472

Fast and precise detection of DNA methylation with tetramethylammonium-filled nanopore

Ejemplares similares