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Cited by 188

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An Efficient RNA-Cleaving DNA Enzyme that Synchronizes Catalysis with Fluorescence Signaling

Mei, Shirley H. J., Liu, Zhongjie, Brennan, John D., Li, Yingfu·2003·Journal of the American Chemical Society

Abstract

DNA enzymes are single-stranded DNA molecules with catalytic capabilities that are isolated from random-sequence DNA libraries by "in vitro selection". This new class of catalytic biomolecules has the potential of being used as unique molecular tools in a variety of innovative applications. Here we describe the creation and characterization of an RNA-cleaving autocatalytic DNA, DEC22-18, that uniquely links chemical catalysis with real-time fluorescence signaling capability in the same molecule. A trans-acting DNA molecule, DET22-18, was also developed from DEC22-18 that behaves as a true enzyme with a k(cat) of approximately 7 min(-1)-a rate constant that is the second largest ever reported for a DNA enzyme. It cleaves a chimeric RNA/DNA substrate at the lone RNA linkage surrounded by a closely spaced fluorophore-quencher pair-a unique structure that permits the synchronization of the chemical cleavage with fluorescence signaling. DET22-18 has a stem-loop structure and can be conjugated with DNA aptamers to form allosteric deoxyribozyme biosensors.

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Authors
Mei, S. H. J., Liu, Z., Brennan, J. D., & Li, Y.
Venue
Journal of the American Chemical Society
Published
2002-12-13
Volume
125
Issue
2
Pages
412-420
ISSN
  • 0002-7863 (ISSN-L)
  • 1520-5126
OpenAlex
W1982237778
Cited by
188
References
0
FWCI
2.05
Type
journal article
Language
en
Open access
closed

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