title: One-pot approach for examining the DNA methylation patterns using an engineered methyl-probe.
abstract: Aberrant DNA methylation is a common observation in various types of human cancers, i.e., breast and lung cancers. Nevertheless, the current DNA methylation detection approaches require bisulfite treatments and are laborious or costly to perform. To address these challenges, we developed a methyl-probe based on the MBD1 protein. Combined with fluorescence correlation spectroscopy, our probe can sensitively detect the existence of DNA methylation at concentrations above 20nM in a one-pot assay. The probe can quantify the total amount of methylated CG dinucleotides above ~20nM, independent of DNA sequence contexts, concentrations (20-1900nM) and methylation levels (5-100%). Our detection platform offers a simple and cheap alternative DNA methylation detection approach.
authors.affiliation: School of Chemical Engineering, Purdue University, 480 Stadium Mall Drive, West Lafayette, IN 47907, USA.
authors.forename: Seong-Eun
authors.initials: SE
authors.lastname: Kim
authors.affiliation: School of Chemical Engineering, Purdue University, 480 Stadium Mall Drive, West Lafayette, IN 47907, USA.
authors.forename: Matthew
authors.initials: M
authors.lastname: Chang
authors.affiliation: School of Chemical Engineering, Purdue University, 480 Stadium Mall Drive, West Lafayette, IN 47907, USA.
authors.forename: Chongli
authors.initials: C
authors.lastname: Yuan
mesh_terms: D020137:Base Pair Mismatch; D001483:Base Sequence; D015374:Biosensing Techniques / Q000295:instrumentation*; D019175:DNA Methylation / Q000235:genetics*; D015342:DNA Probes / Q000737:chemistry* / Q000235:genetics*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005818:Genetic Engineering / Q000379:methods*; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA / Q000295:instrumentation*; D013050:Spectrometry, Fluorescence / Q000295:instrumentation*
chemical_list: D015342:DNA Probes
keywords: DNA methylation; Fluorescence correlation spectroscopy; Methyl-binding domain