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...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2017-03-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-017-00317-2 |
| _version_ | 1818749351813447680 |
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| 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 |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-12-18T04:02:24Z |
| format | Article |
| id | doaj.art-d9155eca816146ab9618e01d64b28e49 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-18T04:02:24Z |
| publishDate | 2017-03-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-d9155eca816146ab9618e01d64b28e492022-12-21T21:21:40ZengNature PortfolioScientific Reports2045-23222017-03-017111010.1038/s41598-017-00317-2Fast and precise detection of DNA methylation with tetramethylammonium-filled nanoporeYing Wang0Yani Zhang1Yanli Guo2Xiao-feng Kang3Key Laboratory of Synthetic and Natural Functional Molecular Chemistry, College of Chemistry & Materials Science, Northwest UniversityCollege of Life Sciences, Northwest UniversityKey Laboratory of Synthetic and Natural Functional Molecular Chemistry, College of Chemistry & Materials Science, Northwest UniversityKey Laboratory of Synthetic and Natural Functional Molecular Chemistry, College of Chemistry & Materials Science, Northwest UniversityAbstract 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.https://doi.org/10.1038/s41598-017-00317-2 |
| spellingShingle | Ying Wang Yani Zhang Yanli Guo Xiao-feng Kang Fast and precise detection of DNA methylation with tetramethylammonium-filled nanopore Scientific Reports |
| title | 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_short | Fast and precise detection of DNA methylation with tetramethylammonium-filled nanopore |
| title_sort | fast and precise detection of dna methylation with tetramethylammonium filled nanopore |
| url | https://doi.org/10.1038/s41598-017-00317-2 |
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