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Issue 2, 2016
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Detection of DNA methyltransferase activity using allosteric molecular beacons

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Abstract

Abnormal DNA methylation patterns caused by altered DNA methyltransferase (MTase) activity are closely associated with cancer. Herein, using DNA adenine methylation methyltransferase (Dam MTase) as a model analyte, we designed an allosteric molecular beacon (aMB) for sensitive detection of Dam MTase activity. When the specific site in an aMB is methylated by Dam MTase, the probe can be cut by the restriction nuclease DpnI to release a fluorophore labeled aptamer specific for streptavidin (SA) which will bind to SA beads to generate highly fluorescent beads for easy signal readout by a microscope or flow cytometer. However, aMBs maintain a hairpin structure without the binding ability to SA beads in the absence of Dam MTase, leading to weakly fluorescent SA beads. Unlike the existing signal amplified assays, our method is simpler and more convenient. The high performance of the aptamer and the easy bead separation process make this probe superior to other methods for the detection of MTase in complex biological systems. Overall, the proposed method with a detection limit of 0.57 U mL−1 for Dam MTase shows great potential for further applications in the detection of other MTases, screening of MTase inhibitors, and early diagnosis of cancer.

Graphical abstract: Detection of DNA methyltransferase activity using allosteric molecular beacons

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Publication details

The article was received on 28 Aug 2015, accepted on 04 Oct 2015 and first published on 05 Oct 2015


Article type: Paper
DOI: 10.1039/C5AN01763G
Author version available: Download Author version (PDF)
Citation: Analyst, 2016,141, 579-584
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    Detection of DNA methyltransferase activity using allosteric molecular beacons

    W. Zhang, X. Zu, Y. Song, Z. Zhu and C. J. Yang, Analyst, 2016, 141, 579
    DOI: 10.1039/C5AN01763G

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