Please use this identifier to cite or link to this item: https://doi.org/10.1038/s41598-018-31197-9
DC FieldValue
dc.titleA novel molecular rotor facilitates detection of p53-DNA interactions using the Fluorescent Intercalator Displacement Assay
dc.contributor.authorGoh, W.L
dc.contributor.authorLee, M.Y
dc.contributor.authorLim, T.X
dc.contributor.authorChua, J.S
dc.contributor.authorBrenner, S
dc.contributor.authorGhadessy, F.J
dc.contributor.authorTeo, Y.N
dc.date.accessioned2020-10-20T03:25:22Z
dc.date.available2020-10-20T03:25:22Z
dc.date.issued2018
dc.identifier.citationGoh, 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. https://doi.org/10.1038/s41598-018-31197-9
dc.identifier.issn20452322
dc.identifier.urihttps://scholarbank.nus.edu.sg/handle/10635/177810
dc.description.abstractWe 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).
dc.rightsAttribution 4.0 International
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.sourceUnpaywall 20201031
dc.subjectDNA
dc.subjectfluorescent dye
dc.subjectintercalating agent
dc.subjectprotein p53
dc.subjectTP53 protein, human
dc.subjectchemistry
dc.subjecthuman
dc.subjectmetabolism
dc.subjectspectrofluorometry
dc.subjectDNA
dc.subjectFluorescent Dyes
dc.subjectHumans
dc.subjectIntercalating Agents
dc.subjectSpectrometry, Fluorescence
dc.subjectTumor Suppressor Protein p53
dc.typeArticle
dc.contributor.departmentDEPT OF MEDICINE
dc.description.doi10.1038/s41598-018-31197-9
dc.description.sourcetitleScientific Reports
dc.description.volume8
dc.description.issue1
dc.description.page12946
Appears in Collections:Staff Publications
Elements

Show simple item record
Files in This Item:
File Description SizeFormatAccess SettingsVersion 
10_1038_s41598-018-31197-9.pdf3.62 MBAdobe PDF

OPEN

NoneView/Download

Google ScholarTM

Check

Altmetric


This item is licensed under a Creative Commons License Creative Commons