Journal
BIOSENSORS & BIOELECTRONICS
Volume 93, Issue -, Pages 293-297Publisher
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2016.08.073
Keywords
Deoxyribonuclease I; RNA/DNA hybrid; DNA-templated silver nanocluster; Graphene oxide; Nanocomposite; Fluorescent enzyme assay
Categories
Funding
- Mid-career Researcher Support Program [2015R1A2A1A01005393]
- Bio-Synergy Research Project through the National Research Foundation - Ministry of Science, ICT and Future Planning (MSIP) of Korea [2014M3A9C4066457]
- Center for BioNano Health-Guard - MSIP of Korea as Global Frontier Project [H-GUARD_2013M3A6B2078964]
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A novel label-free system for the sensitive fluorescent detection of deoxyribonuclease I (DNase I) activity has been developed by utilizing DNA-templated silver nanocluster/graphene oxide (DNA-AgNC/GO) nanocomposite. AgNC is first synthesized around C-rich template DNA and the resulting DNA-AgNC binds to GO through the interaction between the extension DNA and GO. The resulting DNA-AgNC/GO would show quite reduced fluorescence signal because the fluorescence from DNA-AgNCs is quenched by GO. In the presence of DNase I, however, it degrades the DNA strand within DNA/RNA hybrid duplex probe employed in this study, consequently releasing RNA which is complementary to the extension DNA. The released free RNA then extracts DNA-AgNC from GO by hybridizing with the extension DNA bound to GO. This process would restore the quenched fluorescence, emitting highly enhanced fluorescence signal. By employing this assay principle, DNase I activity was reliably identified with a detection limit of 0.10 U/ml which is lower than those from previous fluorescence-based methods. Finally, the practical capability of this assay system was successfully demonstrated by its use to determine DNase I activity in bovine urine. (C) 2016 Elsevier B.V. All rights reserved.
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