Issue 18, 2017

A novel label-free fluorescent molecular beacon for the detection of 3′–5′ exonuclease enzymatic activity using DNA-templated copper nanoclusters

Abstract

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.

Graphical abstract: A novel label-free fluorescent molecular beacon for the detection of 3′–5′ exonuclease enzymatic activity using DNA-templated copper nanoclusters

Supplementary files

Article information

Article type
Paper
Submitted
20 May 2017
Accepted
27 Jul 2017
First published
28 Jul 2017

New J. Chem., 2017,41, 9718-9723

A novel label-free fluorescent molecular beacon for the detection of 3′–5′ exonuclease enzymatic activity using DNA-templated copper nanoclusters

J. Ge, Z. Dong, D. Bai, L. Zhang, Y. Hu, D. Ji and Z. Li, New J. Chem., 2017, 41, 9718 DOI: 10.1039/C7NJ01761H

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