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Issue 18, 2015
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NanoCluster Beacons as reporter probes in rolling circle enhanced enzyme activity detection

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Abstract

As a newly developed assay for the detection of endogenous enzyme activity at the single-catalytic-event level, Rolling Circle Enhanced Enzyme Activity Detection (REEAD) has been used to measure enzyme activity in both single human cells and malaria-causing parasites, Plasmodium sp. Current REEAD assays rely on organic dye-tagged linear DNA probes to report the rolling circle amplification products (RCPs), the cost of which may hinder the widespread use of REEAD. Here we show that a new class of activatable probes, NanoCluster Beacons (NCBs), can simplify the REEAD assays. Easily prepared without any need for purification and capable of large fluorescence enhancement upon hybridization, NCBs are cost-effective and sensitive. Compared to conventional fluorescent probes, NCBs are also more photostable. As demonstrated in reporting the human topoisomerases I (hTopI) cleavage-ligation reaction, the proposed NCBs suggest a read-out format attractive for future REEAD-based diagnostics.

Graphical abstract: NanoCluster Beacons as reporter probes in rolling circle enhanced enzyme activity detection

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Supplementary files

Article information


Submitted
16 Mar 2015
Accepted
08 Apr 2015
First published
14 Apr 2015

Nanoscale, 2015,7, 8332-8337
Article type
Communication

NanoCluster Beacons as reporter probes in rolling circle enhanced enzyme activity detection

S. Juul, J. M. Obliosca, C. Liu, Y. Liu, Y. Chen, D. M. Imphean, B. R. Knudsen, Y. Ho, K. W. Leong and H. Yeh, Nanoscale, 2015, 7, 8332
DOI: 10.1039/C5NR01705J

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