Electrochemical Assay for the Signal-On Detection of Human DNA Methyltransferase Activity
- Creators
- Muren, Natalie B.
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Barton, Jacqueline K.
Abstract
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.
Additional Information
© 2013 American Chemical Society. Received: August 19, 2013; Published: October 28, 2013. This work was supported by NIH GM61077. We also thank the Parsons Foundation for a fellowship to NBM. We thank C. Pheeney and P. Bartels for assistance with multiplexed chip fabrication. Multiplexed chip fabrication was completed in the Kavli Nanoscience Institute at Caltech.Attached Files
Accepted Version - nihms532330.pdf
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Additional details
- PMCID
- PMC3899840
- Eprint ID
- 42852
- Resolver ID
- CaltechAUTHORS:20131205-093720227
- GM61077
- NIH
- Ralph M. Parsons Foundation
- Created
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2013-12-05Created from EPrint's datestamp field
- Updated
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2021-11-10Created from EPrint's last_modified field
- Caltech groups
- Kavli Nanoscience Institute