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- Title
A methylation-stimulated DNA machine: an autonomous isothermal route to methyltransferase activity and inhibition analysis.
- Authors
Changfeng Zhu; Yanqin Wen; Hongzhen Peng; Yitao Long; Yao He; Qing Huang; Di Li; Chunhai Fan
- Abstract
The operation of DNA nanomachines is generally triggered by either conformational changes of DNA nanostructure or external environmental stimuli. In the present study, we demonstrate an alternative driving force, DNA methylation, to stimulate DNA machine operation. DNA methylation changes neither DNA sequence and conformation nor external environment, however, blocks its cleavage by corresponding methylation-sensitive restriction endonuclease. We thus designed a strand displacement amplification DNA machine, which could be stimulated upon DNA methylation and then autonomously generates accumulated amounts of peroxidase-mimicking DNAzyme signaling machine products in an isothermal manner. The machine product DNAzyme could catalyze the HO-mediated oxidation of 2,2′-azino-bis(3-ethylbenzo thiazoline-6-sulfonic acid) (ABTS) to a colored product ABTS. This methylation-stimulated DNA machine was further used as a colorimetric assay for analysis of methyltransferases activities and screening of methylation inhibitors. As compared with classical methylation assay, this facile isothermal DNA machine avoids the introduction of methylation-specific polymerase chain reaction and radioactive labels, which might be employed as an effective tool for DNA methylation analysis.
- Subjects
DNA; METHYLATION; METHYLTRANSFERASES; ENZYMES; NANOSTRUCTURES
- Publication
Analytical & Bioanalytical Chemistry, 2011, Vol 399, Issue 10, p3459
- ISSN
1618-2642
- Publication type
Academic Journal
- DOI
10.1007/s00216-010-4137-2