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A novel label-free fluorescent molecular beacon for the detection of 3′–5′ exonuclease enzymatic activity using DNA-templated copper nanoclusters
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 ex...
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Published in: | New journal of chemistry 2017, Vol.41 (18), p.9718-9723 |
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Main Authors: | , , , , , , |
Format: | Article |
Language: | English |
Citations: | Items that this one cites Items that cite this one |
Online Access: | Get full text |
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Summary: | 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. |
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ISSN: | 1144-0546 1369-9261 |
DOI: | 10.1039/C7NJ01761H |