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Thioflavin T Specifically Brightening “Guanine Island” in Duplex-DNA: A Novel Fluorescent Probe for Single-Nucleotide Mutation

Abstract

Here, we found that Thioflavin T (ThT) could specifically bind with a G-GGG unit (named as “Guanine Island”) in double stranded DNA (ds-DNA). Through stacking with G base via hydrogen bonds, the rotation of ThT was restricted, and concurrently its planarization was enforced. Such a binding mode produced a significantly enhanced ThT fluorescence. Based on the discovery, with ThT as a lighting up probe for “Guanine Island” in ds-DNA, the fluorescent detection of single-nucleotide mutation (C mutation) was investigated. Upon C base in target DNA mutating to G, A or T, and further hybridizing with a probe DNA containing GGG unit at the mutated point, ds-DNA including G-GGG, A-GGG or T-GGG island was formed, respectively. After binding with ThT, C mutation was clearly recognized. With C mutating to G as an example, the detection limit was as low as 3 nM. Importantly, the developed assay could be applied to recognize C mutating to G occurred in a DNA sequence related with dilated cardiomyopathy in diluted human serum. As a sensing unit (“Guanine Island” in ds-DNA lighting up ThT), it is expected to be applied in various biological or environmental systems.

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Supplementary files

Publication details

The article was received on 15 Dec 2018, accepted on 12 Feb 2019 and first published on 13 Feb 2019


Article type: Paper
DOI: 10.1039/C8AN02430H
Citation: Analyst, 2019, Accepted Manuscript

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    Thioflavin T Specifically Brightening “Guanine Island” in Duplex-DNA: A Novel Fluorescent Probe for Single-Nucleotide Mutation

    X. Li, W. Zhou, Z. Yu, G. Ma, T. Jin, Y. Li and L. Fan, Analyst, 2019, Accepted Manuscript , DOI: 10.1039/C8AN02430H

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