Journal
SCIENTIFIC REPORTS
Volume 3, Issue -, Pages -Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/srep01389
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Funding
- National Institutes of Health [R21 CA155863, NIDDK R01 053884]
- Midwestern Cancer Nanotechnology Training Center at UIUC [NIH-NCI R25 CA154015]
- NIH [R21 CA155863, P41-RR005969]
- Mayo Clinic/University of Illinois Strategic Alliance for Technology-Based Healthcare
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Epigenetic modifications in eukaryotic genomes occur primarily in the form of 5-methylcytosine (5mC). These modifications are heavily involved in transcriptional repression, gene regulation, development and the progression of diseases including cancer. We report a new single-molecule assay for the detection of DNA methylation using solid-state nanopores. Methylation is detected by selectively labeling methylation sites with MBD1 (MBD-1x) proteins, the complex inducing a 3 fold increase in ionic blockage current relative to unmethylated DNA. Furthermore, the discrimination of methylated and unmethylated DNA is demonstrated in the presence of only a single bound protein, thereby giving a resolution of a single methylated CpG dinucleotide. The extent of methylation of a target molecule could also be coarsely quantified using this novel approach. This nanopore-based methylation sensitive assay circumvents the need for bisulfite conversion, fluorescent labeling, and PCR and could therefore prove very useful in studying the role of epigenetics in human disease.
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