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A label-free and enzyme-free signal amplification strategy for a sensitive RNase H activity assay

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Abstract

We herein describe a label-free and enzyme-free signal amplification strategy for the sensitive determination of ribonuclease H (RNase H) activity, which relies on the target-triggered catalytic hairpin assembly (CHA) in conjunction with a G-quadruplex specific fluorescent binder, N-methyl mesoporphyrin IX (NMM). In the absence of RNase H, the RNA/DNA duplex serving as a substrate for RNase H cannot initiate the execution of CHA that produces G-quadruplexes; so NMM shows a low fluorescence signal. In contrast, the presence of RNase H that degrades RNA in the RNA/DNA duplex releases DNA designed to function as the catalyst for CHA. This consequently promotes the efficient CHA and generates a large number of G-quadruplexes with a significantly enhanced fluorescence signal from NMM. Based on this label-free and enzyme-free signal amplification strategy, we successfully determined the RNase H activity with a detection limit of 0.037 U mL−1 and screened potential RNase H inhibitors. Our results suggest that the developed system is a promising platform for a cost-effective, sensitive enzyme activity assay and inhibitor screening.

Graphical abstract: A label-free and enzyme-free signal amplification strategy for a sensitive RNase H activity assay

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Publication details

The article was received on 07 Jun 2017, accepted on 11 Sep 2017 and first published on 14 Sep 2017


Article type: Paper
DOI: 10.1039/C7NR04060A
Citation: Nanoscale, 2017, Advance Article
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    A label-free and enzyme-free signal amplification strategy for a sensitive RNase H activity assay

    C. Y. Lee, H. Jang, K. S. Park and H. G. Park, Nanoscale, 2017, Advance Article , DOI: 10.1039/C7NR04060A

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