Visible Fluorescent Light-up Probe for DNA Three-Way Junctions Provides Host−Guest Biosensing Applications

dc.contributor.authorVan Riesen, Abigail J.
dc.contributor.authorLe, Jennifer
dc.contributor.authorSlavkovic, Sladjana
dc.contributor.authorChurcher, Zachary R.
dc.contributor.authorShoara, Aron A.
dc.contributor.authorJohnson, Philip E.
dc.contributor.authorManderville, Richard A.
dc.date.accessioned2023-05-10T17:04:58Z
dc.date.available2023-05-10T17:04:58Z
dc.date.issued2021-08-18
dc.description.abstractDNA three-way junctions (3WJs) consist of a Y-shaped hydrophobic branch point connecting three double-stranded stems and are viewed as druggable targets for cancer treatment. They are also important building blocks for the construction of DNA nanostructures and serve as recognition elements for DNA aptasensors for a wide variety of diagnostic applications. However, visible fluorescent light-up probes for specific staining of DNA 3WJs are currently lacking. Herein, we report that a merocyanine containing the N-methylbenzothiazolium (Btz) acceptor vinyl linked to a 2-fluorophenolic (FPhO) donor (FPhOBtz) serves as a universal fluorescent turn-on dye for DNA 3WJs. Our evidence is based on a multifaceted approach to define the specificity and affinity of FPhOBtz for 3WJ DNA aptamers; the cocaine binding aptamer MN4, the cholic acid binding aptamer (CABA), and four steroid aptamers (DOGS.1, DISS.1, BES.1, DCAS.1). FPhOBtz exhibits impressive turn-on (up to 730-fold) fluorescence at 580 nm upon aptamer binding with low micromolar affinity. Direct FPhOBtz displacement from the 3WJ binding domain through competitive alkaloid and steroid binding provides immediate fluorescent read out for host−guest detection strategies in human blood serum in the low micromolar regime. Our results present the first visible light-up fluorescent probe for DNA 3WJ detection strategies.en_US
dc.identifier.citationACS Applied Bio Materials 4 (2021): 6732−6741en_US
dc.identifier.urihttps://doi.org/10.1021/acsabm.1c00431en_US
dc.identifier.urihttp://hdl.handle.net/10315/41139
dc.language.isoenen_US
dc.publisherAmerican Chemical Societyen_US
dc.rightsAttribution-NonCommercial-NoDerivatives 4.0 International*
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/*
dc.subjectDNA three-way junctions, fluorescent probe, merocyanine, host−guest detection, cocaine, quinineen_US
dc.titleVisible Fluorescent Light-up Probe for DNA Three-Way Junctions Provides Host−Guest Biosensing Applicationsen_US
dc.typeArticleen_US

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