Detection and Quantification of Methylation in DNA using Solid-State Nanopores

作者：Shim Jiwook; Humphreys Gwendolyn I; Venkatesan Bala Murali; Munz Jan Marie; Zou Xueqing; Sathe Chaitanya; Schulten Klaus; Kosari Farhad; Nardulli Ann M^*; Vasmatzis George; Bashir Rashid

来源：Scientific Reports, 2013, 3(1): 1389.

DOI：10.1038/srep01389

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.

• 出版日期2013-3-11