Electrochemical DNA sensor-based strategy for sensitive detection of DNA demethylation and DNA demethylase activity
| Autor: | Mengxing Fan, Yin Yang, Hui Zhang, Qingming Shen |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
Surface Properties
Biosensing Techniques 010402 general chemistry 01 natural sciences Biochemistry Analytical Chemistry Endonuclease chemistry.chemical_compound Demethylase activity Environmental Chemistry DNA Cleavage DNA Modification Methylases Electrodes Spectroscopy Demethylation chemistry.chemical_classification biology Chemistry 010401 analytical chemistry DNA Electrochemical Techniques 0104 chemical sciences DNA demethylase activity DNA Demethylation DNA demethylation Enzyme biology.protein Demethylase Gold |
| Zdroj: | Analytica chimica acta. 934 |
| ISSN: | 1873-4324 |
| Popis: | DNA demethylation and demethylase activity play important roles in DNA self-repair, and their detection is key to early diagnosis of fatal diseases. Herein, a facile electrochemical DNA (E-DNA) sensor was developed for the sensitive detection of DNA demethylation and demethylase activity based on an enzyme cleavage strategy. The thiol modified hemi-methylated hairpin probe DNA (pDNA) was self-assembled on a Au electrode surface through the formation of Au S bonds. The hemi-methylated pDNA served as the substrate of DNA demethylase (using methyl-CpG-binding domain protein 2 (MBD2) as an example). Following demethylation, the hairpin stem was then recognized and cleaved by BstUI endonuclease. The ferrocene carboxylic acid (FcA)-tagged pDNA strands were released into the buffer solution from the electrode surface, resulting in a significant decrease of electrochemical signal and providing a means to observe DNA demethylation. The activity of DNA demethylase was analyzed in the concentration ranging from 0.5 to 500 ng mL −1 with a limit of detection as low as 0.17 ng mL −1 . With high specificity and sensitivity, rapid response, and low cost, this simple E-DNA sensor provides a unique platform for the sensitive detection of DNA demethylation, DNA demethylase activity, and related molecular diagnostics and drug screening. |
| Databáze: | OpenAIRE |
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