Simple and convenient G-quadruplex-based turn-on fluorescence assay for 3' → 5' exonuclease activity.
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.
Chung-Hang
CH
Leung
Daniel Shiu-Hin
DS
Chan
Bradley Yat-Wah
BY
Man
Chuan-Jen
CJ
Wang
Wing
W
Lam
Yung-Chi
YC
Cheng
Wang-Fun
WF
Fong
Wen-Luan Wendy
WL
Hsiao
Dik-Lung
DL
Ma
D002942:Circular Dichroism; D005090:Exodeoxyribonucleases / Q000378:metabolism; D005092:Exonucleases / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000737:chemistry*; D054856:G-Quadruplexes; D005840:Gentian Violet / Q000737:chemistry*; D006801:Humans; D009841:Oligonucleotides / Q000737:chemistry / Q000378:metabolism; D010750:Phosphoproteins / Q000378:metabolism; D013050:Spectrometry, Fluorescence / Q000379:methods*
D005456:Fluorescent Dyes; D009841:Oligonucleotides; D010750:Phosphoproteins; D005090:Exodeoxyribonucleases; D005092:Exonucleases; C020133:exodeoxyribonuclease III; C429526:three prime repair exonuclease 1; D005840:Gentian Violet