Thioflavin T as a fluorescence light-up probe for G4 formation |
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Authors: | Amandine Renaud de la Faverie Aurore Guédin Amina Bedrat Liliya A. Yatsunyk Jean-Louis Mergny |
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Affiliation: | 1.ARNA Laboratory, University of Bordeaux, F-33000 Bordeaux, France, 2.INSERM U869, IECB, F-33600 Pessac, France and 3.Department of Chemistry and Biochemistry, Swarthmore College, Swarthmore, PA 19081, USA |
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Abstract: | Thioflavin T (ThT) becomes fluorescent in the presence of the G-quadruplex structure such as that formed by the human telomeric motif. In this report, we extend and generalize these observations and show that this dye may be used as a convenient and specific quadruplex probe. In the presence of most, but not all, G4-forming sequences, we observed a large increase in ThT fluorescence emission, whereas the presence of control duplexes and single strands had a more limited effect on emission. This differential behavior allowed us to design a high-throughput assay to detect G4 formation. Hundreds of different oligonucleotides may be tested in parallel for G4 formation with a simple fluorescence plate reader. We applied this technique to a family of aptamers not previously recognized as G4-forming sequences and demonstrated that ThT fluorescence signal may be used to predict G4 formation. |
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