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Title: A novel molecular rotor facilitates detection of p53-DNA interactions using the Fluorescent Intercalator Displacement Assay
Authors: Goh, W.L
Lee, M.Y
Lim, T.X
Chua, J.S 
Brenner, S 
Ghadessy, F.J
Teo, Y.N
Keywords: DNA
fluorescent dye
intercalating agent
protein p53
TP53 protein, human
Fluorescent Dyes
Intercalating Agents
Spectrometry, Fluorescence
Tumor Suppressor Protein p53
Issue Date: 2018
Citation: Goh, W.L, Lee, M.Y, Lim, T.X, Chua, J.S, Brenner, S, Ghadessy, F.J, Teo, Y.N (2018). A novel molecular rotor facilitates detection of p53-DNA interactions using the Fluorescent Intercalator Displacement Assay. Scientific Reports 8 (1) : 12946. ScholarBank@NUS Repository.
Rights: Attribution 4.0 International
Abstract: We have investigated the use of fluorescent molecular rotors as probes for detection of p53 binding to DNA. These are a class of fluorophores that undergo twisted intramolecular charge transfer (TICT). They are non-fluorescent in a freely rotating conformation and experience a fluorescence increase when restricted in the planar conformation. We hypothesized that intercalation of a molecular rotor between DNA base pairs would result in a fluorescence turn-on signal. Upon displacement by a DNA binding protein, measurable loss of signal would facilitate use of the molecular rotor in the fluorescent intercalator displacement (FID) assay. A panel of probes was interrogated using the well-established p53 model system across various DNA response elements. A novel, readily synthesizable molecular rotor incorporating an acridine orange DNA intercalating group (AO-R) outperformed other conventional dyes in the FID assay. It enabled relative measurement of p53 sequence-specific DNA interactions and study of the dominant-negative effects of cancer-associated p53 mutants. In a further application, AO-R also proved useful for staining apoptotic cells in live zebrafish embryos. © 2018, The Author(s).
Source Title: Scientific Reports
ISSN: 20452322
DOI: 10.1038/s41598-018-31197-9
Rights: Attribution 4.0 International
Appears in Collections:Staff Publications

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