Aptamer-mediated universal enzyme assay based on target-triggered DNA polymerase activity

We herein describe an innovative method for a universal fluorescence turn-on enzyme assay, which relies on the target enzyme-triggered DNA polymerase activity. In the first target recognition step, the target enzyme is designed to destabilize detection probe derived from an aptamer specific to DNA polymerase containing the overhang sequence and the complementary blocker DNA, which consequently leads to the recovery of DNA polymerase activity inhibited by the detection probe. This target-triggered polymerase activity is monitored in the second signal transduction step based on primer extension reaction coupled with TaqMan probe. Utilizing this design principle, we have successfully detected the activities of two model enzymes, exonuclease I and uracil DNA glycosylase with high sensitivity and selectivity. Since this strategy is composed of separated target recognition and signal transduction modules, it could be universally employed for the sensitive determination of numerous different target enzymes by simply redesigning the overhang sequence of detection probe, while keeping TaqMan probe-based signal transduction module as a universal signaling tool.
Publisher
ELSEVIER ADVANCED TECHNOLOGY
Issue Date
2017-02
Language
English
Keywords

EXCISION-REPAIR; MOLECULAR BEACONS; GRAPHENE OXIDE; RESTRICTION ENDONUCLEASES; SENSITIVE DETECTION; NUCLEIC-ACID; I ACTIVITY; INHIBITORS; PROBE; SYSTEM

Citation

BIOSENSORS & BIOELECTRONICS, v.88, pp.48 - 54

ISSN
0956-5663
DOI
10.1016/j.bios.2016.07.038
URI
http://hdl.handle.net/10203/214784
Appears in Collection
CBE-Journal Papers(저널논문)
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