A novel label-free fluorescent molecular beacon for the detection of 3′–5′ exonuclease enzymatic activity using DNA-templated copper nanoclusters
| Autor: | Danyang Ji, Dong-Mei Bai, Jia Ge, Zhaohui Li, Zhen-Zhen Dong, Lin Zhang, Yalei Hu |
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| Rok vydání: | 2017 |
| Předmět: |
Exonuclease III
Exonuclease Nuclease biology Chemistry Oligonucleotide 02 engineering and technology General Chemistry 010402 general chemistry 021001 nanoscience & nanotechnology 01 natural sciences Fluorescence Catalysis 0104 chemical sciences chemistry.chemical_compound Biochemistry Molecular beacon Materials Chemistry biology.protein 3'-5' Exonuclease Biophysics 0210 nano-technology DNA |
| Zdroj: | New Journal of Chemistry. 41:9718-9723 |
| ISSN: | 1369-9261 1144-0546 |
| DOI: | 10.1039/c7nj01761h |
| Popis: | The 3′–5′ exonuclease plays key roles in maintaining genome stability; therefore, the detection of 3′–5′ exonuclease activity is very important for disease diagnosis and drug development. In this study, we present a novel highly sensitive and label-free fluorescent assay to detect the activity of exonuclease III (Exo III) using molecular beacon DNA with a polythymine (T) loop as a template for the formation of fluorescent copper nanoparticles (CuNPs). Upon the addition of Exo III, the molecular beacon probe was digested into mono- or short oligonucleotides by the exonucleases and failed to form fluorescent CuNPs. As a result, the fluorescence intensity of CuNPs decreased. This method exhibited a very low detection limit of 0.02 U mL−1 for Exo III. The designed molecular beacon probe can be a potential label-free strategy for the detection of other nuclease activity. |
| Databáze: | OpenAIRE |
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