Abstract |
The level of DNA methylation has been reported to be closely associated with various carcinomas and is dynamically regulated by several demethylases. However, current demethylase detection methods are mainly antibody-based, which detect the demethylase concentrations, while the actual numbers of catalytically active demethylase remain unknown. Thus, we have developed an activity assay based on epigenetically modified DNAzymes (EMOzymes), and CRISPR/Cas12a facilitated cascade signal amplification. We have ultrasensitively quantitated the activity of O6-methylguanine-DNA-methyltransferase (MGMT), a key demethylase that contributes to the chemoresistance to alkylation agents in cancer therapy, with an estimated limit of detection of 0.054 nM. This approach opens a new avenue for sensitively profiling the activity of many disease-related demethylases.
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Authors | Juan Huang, Jiao Wang, Zhenkun Wu, Jianjun He, Jian-Hui Jiang |
Journal | Biosensors & bioelectronics
(Biosens Bioelectron)
Vol. 207
Pg. 114186
(Jul 01 2022)
ISSN: 1873-4235 [Electronic] England |
PMID | 35316758
(Publication Type: Journal Article)
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Copyright | Copyright © 2022 Elsevier B.V. All rights reserved. |
Chemical References |
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Topics |
- Biosensing Techniques
(methods)
- DNA Methylation
- DNA, Catalytic
(metabolism)
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