title: A supercharged fluorescent protein as a versatile probe for homogeneous DNA detection and methylation analysis.
abstract: Supercharged proteins are a new class of functional proteins with exceptional stability and potent ability to deliver bio-macromolecules into cells. As a proof-of-principle, a novel application of supercharged proteins as a versatile biosensing platform for nucleic acid detection and epigenetics analysis is presented. Taking supercharged green fluorescent protein (ScGFP) as the signal reporter, a simple turn-on homogenous method for DNA detection has been developed based on the polyionic nanoscale complex of ScGFP/DNA and toehold strand displacement. This assay shows high sensitivity and potent ability to detect single-base mismatch. Furthermore, combined with bisulfite conversion, this ScGFP-based assay was further applied to analyze site-specific DNA methylation status of genomic DNA extracted from real human colon carcinoma tissue sample with ultrahigh sensitivity (4 amol methylated DNA).
authors: State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, ChangSha 410082 (P.R. China).
authors: Chunyang
authors: C
authors: Lei
authors: Yan
authors: Y
authors: Huang
authors: Zhou
authors: Z
authors: Nie
authors: Jun
authors: J
authors: Hu
authors: Lijun
authors: L
authors: Li
authors: Guoyan
authors: G
authors: Lu
authors: Yitao
authors: Y
authors: Han
authors: Shouzhuo
authors: S
authors: Yao
mesh_terms: D004247:DNA / Q000378:metabolism*; D019175:DNA Methylation; D006801:Humans; D008856:Microscopy, Fluorescence; D015394:Molecular Structure; D011506:Proteins
chemical_list: D011506:Proteins; D004247:DNA
keywords: DNA; DNA methylation; bioanalysis; supercharged protein; toehold strand displacement