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Silver ion-stabilized DNA triplexes for completely enzyme-free and sensitive fluorescence detection of transcription factors via catalytic hairpin assembly amplification

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Abstract

Transcription factors play important roles in gene regulation and have been identified as promising biomarkers for disease diagnosis. On the basis of a new Ag+-stabilized DNA triplex probe and catalytic hairpin assembly (CHA) signal amplification, we have established a completely enzyme-free and sensitive method for simple fluorescence detection of NF-κB p50 (nuclear factor-kappa B), a transcription factor. We found that the employment of Ag+ to stabilize the DNA triplex structure could effectively reduce the background noise. The association of the target NF-κB p50 with the recognition hairpin in the Ag+-stabilized DNA triplex leads to the release of a single stranded DNA, which is used as the trigger to initiate subsequent CHA between a fluorescently quenched signal hairpin and the recognition hairpin in the triplex DNA structure, thereby resulting in drastically amplified fluorescence recovery for sensitive detection of NF-κB p50. This assay method shows a dynamic concentration range of 5 to 150 pM and a detection limit of 1.5 pM for the detection of NF-κB p50. Besides, the presence of the target molecules can also be selectively discriminated from other non-specific proteins. Moreover, the presence of low concentrations of NF-κB p50 in human serum samples could be monitored with this approach. With the successful demonstration for NF-κB p50, such a method can be potentially extended to detecting other transcription factors in a convenient and sensitive manner without using any enzymes for signal amplification.

Graphical abstract: Silver ion-stabilized DNA triplexes for completely enzyme-free and sensitive fluorescence detection of transcription factors via catalytic hairpin assembly amplification

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Publication details

The article was received on 19 Nov 2018, accepted on 24 Dec 2018 and first published on 24 Dec 2018


Article type: Paper
DOI: 10.1039/C8TB03042A
Citation: J. Mater. Chem. B, 2019, Advance Article
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    Silver ion-stabilized DNA triplexes for completely enzyme-free and sensitive fluorescence detection of transcription factors via catalytic hairpin assembly amplification

    Y. Qin, D. Li, R. Yuan and Y. Xiang, J. Mater. Chem. B, 2019, Advance Article , DOI: 10.1039/C8TB03042A

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