Colorimetric detection of uracil DNA glycosylase activity based on target-activated toehold-mediated strand displacement circuit = 표적 활성 염기가닥 치환 회로 기반의 uracil DNA glycosylase 비색 검출법

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Herein, we describe a novel enzyme-free and label-free strategy for colorimetric assay of uracil DNA glycosylase (UDG) activity, which relies on a target-activated toehold-mediated strand displacement (TMSD) circuit. The strategy employs a detection probe composed of an uracil-containing strand and a catalyst strand (CS) designed to trigger the TMSD circuit. Thus, in the presence of UDG which cleaves uracil bases and destabilizes the detection probe, CS is released to promote the TMSD reaction. This leads to the liberation of a G-quadruplex DNAzyme strand (GS) with the peroxidase mimicking activity, which is initially caged by a blocker strand. Notably, a fuel strand is supplemented to promote another cycle of TMSD reaction by recycling the CS, which activates a large number of GSs. As a consequence, a distinct colorimetric signal is generated from the oxidation of ABTS by GSs. With this strategy, we selectively determined the UDG activity down to 0.006 U/ml, and also identified its presence with the naked eye. Finally, the practical applicability of this assay was successfully demonstrated by reliably determining UDG activities in human serum.
Advisors
Park, Hyun Gyuresearcher박현규researcher
Description
한국과학기술원 :생명화학공학과,
Publisher
한국과학기술원
Issue Date
2019
Identifier
325007
Language
eng
Description

학위논문(석사) - 한국과학기술원 : 생명화학공학과, 2019.2,[ⅳ, 28 p. :]

Keywords

Uracil DNA glycosylase▼atoehold-mediated strand displacement circuit▼aG-quadruplex DNAzyme▼aperoxidase mimicking activity▼acolorimetric assays; Uracil DNA glycosylase▼a염기가닥 치환 회로▼aG-사중구조 DNA 기반 효소▼aPeroxidase mimicking activity▼a비색 검출

URI
http://hdl.handle.net/10203/266329
Link
http://library.kaist.ac.kr/search/detail/view.do?bibCtrlNo=843225&flag=dissertation
Appears in Collection
CBE-Theses_Master(석사논문)
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