期刊
NANOSCALE
卷 9, 期 42, 页码 16149-16153出版社
ROYAL SOC CHEMISTRY
DOI: 10.1039/c7nr04060a
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资金
- Mid-career Researcher Support Program through the National Research Foundation (NRF) - Ministry of Science, ICT and Future Planning (MSIP) of Korea [2015R1A2A1A01005393]
- Center for BioNano Health-Guard
- MSIP of Korea as a Global Frontier Project [H-GUARD_2013M3A6B2078964]
- NRF grant
- MSIP of Korea [2017R1C1B5017724]
- National Research Foundation of Korea [2015R1A2A1A01005393] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
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.
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