Label-free fluorescence strategy for methyltransferase activity assay based on poly-thymine copper nanoclusters engineered by terminal deoxynucleotidyl transferase
| Autor: | Rui-Hong Deng, Bao-Zhu Chi, Zhimei Li, Yu-Chuan Feng, Ting Pi, Ziyi Xia, Kefang Yang, Xiang-Juan Zheng |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
Methyltransferase
Biosensing Techniques 02 engineering and technology 010402 general chemistry 01 natural sciences Fluorescence Analytical Chemistry chemistry.chemical_compound DNA Nucleotidylexotransferase Instrumentation Spectroscopy Fluorescent Dyes Label free Detection limit Copper nanoclusters 021001 nanoscience & nanotechnology Atomic and Molecular Physics and Optics 0104 chemical sciences Thymine chemistry Terminal deoxynucleotidyl transferase Biochemistry DNA methylation 0210 nano-technology Copper |
| Zdroj: | Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy. 260:119924 |
| ISSN: | 1386-1425 |
| DOI: | 10.1016/j.saa.2021.119924 |
| Popis: | The assay of detecting DNA methyltransferase activity has been identified as one of the central challenges in cancer diagnostics as DNA methylation is closely related to the diagnosis and treatment of tumors. In this study, a label-free fluorescence probe based on poly-thymine copper nanoclusters engineered by terminal deoxynucleotidyl transferase is proposed for methyltransferase activity assay. Taking advantage of the efficient polymerization extension reaction catalyzed by terminal deoxynucleotidyl transferase and the copper nanoclusters with large Stokes shift instead of labeling fluorescent dyes, the strategy exhibits a broader linear scope from 1 to 300 U mL−1 with a detection limit of 0.176 U mL−1. The economical method is specificity for M.SssI and also provides a convenient and high-throughput platform for screening the DNA methylation inhibitors, which displays a great potential for the practical applications of the drug development and clinical cancer diagnosis in the future. |
| Databáze: | OpenAIRE |
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