Electrochemical assay for the signal-on detection of human DNA methyltransferase activity

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[1] Muren, Natalie B.
[2] Barton, Jacqueline K.
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Barton, J.K. (jkbarton@caltech.edu) | 1600年 / American Chemical Society卷 / 135期
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Strategies to detect human DNA methyltransferases are needed; given that aberrant methylation by these enzymes is associated with cancer initiation and progression. Here we describe a nonradioactive; antibody-free; electrochemical assay in which methyltransferase activity on DNA-modified electrodes confers protection from restriction for signal-on detection. We implement this assay with a multiplexed chip platform and show robust detection of both bacterial (SssI) and human (Dnmt1) methyltransferase activity. Essential to work with human methyltransferases; our unique assay design allows activity measurements on both unmethylated and hemimethylated DNA substrates. We validate this assay by comparison with a conventional radioactive method. The advantages of electrochemistry over radioactivity and fluorescence make this assay an accessible and promising new approach for the sensitive; label-free detection of human methyltransferase activity. © 2013 American Chemical Society;
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