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
Main Authors: Ge, Jia, Dong, Zhen-Zhen, Bai, Dong-Mei, Zhang, Lin, Hu, Ya-Lei, Ji, Dan-Yang, Li, Zhao-Hui
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container_end_page 9723
container_issue 18
container_start_page 9718
container_title New journal of chemistry
container_volume 41
creator Ge, Jia
Dong, Zhen-Zhen
Bai, Dong-Mei
Zhang, Lin
Hu, Ya-Lei
Ji, Dan-Yang
Li, Zhao-Hui
description 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.
doi_str_mv 10.1039/C7NJ01761H
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title A novel label-free fluorescent molecular beacon for the detection of 3′–5′ exonuclease enzymatic activity using DNA-templated copper nanoclusters
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