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Simple and Convenient G-Quadruplex-Based Turn-On Fluorescence Assay for 3′ → 5′ Exonuclease Activity

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journal contribution
posted on 15.01.2011, 00:00 by Chung-Hang Leung, Daniel Shiu-Hin Chan, Bradley Yat-Wah Man, Chuan-Jen Wang, Wing Lam, Yung-Chi Cheng, Wang-Fun Fong, Wen-Luan Wendy Hsiao, Dik-Lung Ma
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.