Abstract
We previously reported a molecular beacon-mRNA (MB-mRNA) strategy for nucleic acid detection/sensing in a cell-free translation system using unmodified RNA as a probe. Here in this presentation, we report that a combination with RNase H activity, which induces an additional process of irreversible cleavage of MB-domain, achieves an improved sequence selectivity (one nucleotide selectivity) and an enhanced sensitivity. This improved system finally enabled visible sensing of target nucleic acid sequence at a single nucleotide resolution under isothermal conditions.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Base Sequence
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Biosensing Techniques*
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Cell-Free System
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Colorimetry
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Fluorescence Resonance Energy Transfer
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Genes, Reporter
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Luciferases / analysis
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Luciferases / genetics
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Luminescent Agents / analysis
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Luminescent Measurements
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Nucleic Acid Hybridization / methods*
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Nucleic Acids / analysis*
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Oligodeoxyribonucleotides / analysis
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Protein Biosynthesis*
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RNA Probes / chemistry*
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Receptors, CCR5 / genetics
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Ribonuclease H*
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beta-Galactosidase / analysis
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beta-Galactosidase / genetics
Substances
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Luminescent Agents
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Nucleic Acids
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Oligodeoxyribonucleotides
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RNA Probes
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Receptors, CCR5
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Luciferases
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Ribonuclease H
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beta-Galactosidase