An Efficient RNA-Cleaving DNA Enzyme that Synchronizes Catalysis with Fluorescence Signaling
- 13 December 2002
- journal article
- Published by American Chemical Society (ACS) in Journal of the American Chemical Society
- Vol. 125 (2), 412-420
- https://doi.org/10.1021/ja0281232
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 kcat of ∼7 min-1a 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 paira 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.Keywords
This publication has 24 references indexed in Scilit:
- Toward an RNaseA Mimic: A DNAzyme with Imidazoles and Cationic AminesJournal of the American Chemical Society, 2002
- Making Catalytic DNAsScience, 2000
- Ferredoxin-Mediated Electrocatalytic Dehalogenation of Haloalkanes by Cytochrome P450camJournal of the American Chemical Society, 2000
- Cofactor‐Assisted Self‐Cleavage in DNA Libraries with a 3′–5′‐Phosphoramidate BondAngewandte Chemie-International Edition, 1997
- DNA enzymesNature Biotechnology, 1997
- In vitro selection of self-cleaving DNAsCell Chemical Biology, 1996
- A DNA metalloenzyme with DNA ligase activityNature, 1995
- A DNA Aptamer That Binds Adenosine and ATPBiochemistry, 1995
- In vitro selection of RNA molecules that bind specific ligandsNature, 1990
- Systematic Evolution of Ligands by Exponential Enrichment: RNA Ligands to Bacteriophage T4 DNA PolymeraseScience, 1990