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Label-free and enzyme-free signal amplification strategy for the sensitive RNase H activity assay

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, RNA/DNA duplex serving as a substrate for RNase H cannot initiate the execution of CHA that produces G-quadruplexes, so NMM shows the low fluorescence signal. In contrast, the presence of RNase H that degrades RNA in 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 the 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 the detection limit of 0.037 U/mL and screened potential RNase H inhibitors. Our results suggest that the developed system is a promising platform for the cost-effective, sensitive enzyme activity assay and inhibitor screening.

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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, Accepted Manuscript
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    Label-free and enzyme-free signal amplification strategy for the sensitive RNase H activity assay

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

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