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.
journal_name
Sci Repjournal_title
Scientific reportsauthors
Goh WL,Lee MY,Lim TX,Chua JS,Brenner S,Ghadessy FJ,Teo YNdoi
10.1038/s41598-018-31197-9subject
Has Abstractpub_date
2018-08-28 00:00:00pages
12946issue
1issn
2045-2322pii
10.1038/s41598-018-31197-9journal_volume
8pub_type
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