Abstract
A selective, oligonucleotide-based, label-free, turn-on fluorescence detection method for 3' → 5' exonuclease activity has been developed using crystal violet as a G-quadruplex-binding probe. The assay is highly simple and rapid, does not require the use of gel-based equipment or radioisotopic labeling, and is amenable to high-throughput and real-time detection. A proof-of-concept of this assay has been demonstrated for prokaryotic Exonuclease III (ExoIII) and human TREX1.
Publication types
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Letter
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Circular Dichroism
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Exodeoxyribonucleases / metabolism
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Exonucleases / metabolism*
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Fluorescent Dyes / chemistry*
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G-Quadruplexes*
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Gentian Violet / chemistry*
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Humans
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Oligonucleotides / chemistry
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Oligonucleotides / metabolism
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Phosphoproteins / metabolism
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Spectrometry, Fluorescence / methods*
Substances
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Fluorescent Dyes
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Oligonucleotides
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Phosphoproteins
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Exodeoxyribonucleases
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Exonucleases
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exodeoxyribonuclease III
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three prime repair exonuclease 1
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Gentian Violet