Journal
MICROCHIMICA ACTA
Volume 185, Issue 10, Pages -Publisher
SPRINGER WIEN
DOI: 10.1007/s00604-018-3036-7
Keywords
Gold nanoparticles; Aptamer; DNA nanotechnology; DNA walker; Molecular machine; Nicking endonuclease; Autonomous movement; Fluorescence resonance energy transfer; Enzymatic cleavage
Categories
Funding
- National Natural Science Foundation of China [21775019, 21475020, 21635004, 81730087]
- Priority Academic Program Development of Jiangsu Higher Education Institutions [2242018K3DN04]
- Key Laboratory of Modern Toxicology of Ministry of Education, Nanjing Medical University [NMUMT201804]
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The authors describe a fluorometric method for improving the determination of the cancer biomarker 8-hydroxy-2'-deoxyguanosine (8-OHdG). A nicking endonuclease (NEase)-powered 3-D DNA nanomachine was constructed by assembling hundreds of carboxyfluorescein-labeled single strand oligonucleotides (acting as signal reporter) and tens of swing arms (acting as single-foot DNA walkers) on a gold nanoparticle (AuNP). The activity of this DNA nanomachine was controlled by introducing the protecting oligonucleotides. In the presence of aptamer against 8-OHdG, the protecting oligonucleotides are removed from the swing arms by toehold-mediated strand displacement reaction. In the next step, detached DNA walker hybridizes to the labelled DNA so that the DNA nanomachine becomes activated. Special sequences of signal reporter in the formed duplex can be recognized and cleaved by NEase. As a result, the DNA walker autonomously and progressively moves along the surface of the AuNP, thereby releasing hundreds of signal reporters and causing a rapid increase in green fluorescence. This 3-D nanomachine is highly efficient because one aptamer can release hundreds of signal reporters. These unique properties allowed for the construction of a DNA nanomachine-based method for sensitively detecting 8-OHdG in concentrations as low as 4 pM. This is three orders of magnitude lower compared to previously reported methods.
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