A label-free fluorescent molecular switch for a DNA hybridization assay utilizing a G-quadruplex-selective auramine O

Xu, Hao; Geng, Fenghua; Wang, Yongxiang; Xu, Maotian; Lai, Xin‐He; Qu, Peng; Zhang, Yintang; Liu, Baohong

doi:10.1039/c5cc02624e

Cited by 26 publications

(25 citation statements)

References 33 publications

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“…It has also been reported that Auramine‐O decays when it is bound to cellulose (Simkovitch and Huppert ) and solid clays (Valandro et al ). The enhancement in fluorescence intensity is similar to a reported case for the mixing of Auramine‐O with htDNA and the amyloid fibril protein (Amdursky and Huppert ; Xu et al ). By keeping C A > 0·1 µ g ml −1 , Auramine‐O can provide more than 60% of the maximum fluorescence intensity within 10 s, which is very reasonable considering the time required for a rapid microbe measurement.…”

Section: Discussionsupporting

confidence: 87%

“…Auramine‐O is a nonspecific dye that is embedded in the cell membrane and bound to cell proteins or enzymes in cells (Ivanov et al ; Amdursky and Huppert ). Auramine‐O has excitation and emission wavelengths at 427–435 nm and 495–510 nm respectively (Riemersma and Alsbach ; Chen ; Ivanov et al ; Amdursky and Huppert ; Simkovitch and Huppert ; Xu et al ). It has been used in various biological studies as a stain for synthetic polycarboxylic acids (Braud ) and yeast S. cerevisiae (Brennan and Schiestl ).…”

Section: Methodsmentioning

confidence: 99%

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Optimal fluorescent‐dye staining time for the real‐time detection of microbes: a study of Saccharomyces cerevisiae

et al. 2020

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Aims To provide information on the time‐dependent behaviour of microbe staining by fluorescent dyes in the order of seconds, which is important in terms of the recent rapid and online techniques for microbe measurements and/or environmental microbe analysis. Methods and Results For combinations of yeast (Saccharomyces cerevisiae) and typical dyes, including DAPI (4′,6‐diamidino‐2‐phenylindole) and Auramine‐O, a suspension of yeast cells in ultrapure water was injected into a dye solution in a micro cuvette placed inside a spectrofluorometer and the fluorescence intensity of the resulting solution was measured at 1 s intervals, starting immediately after the mixing and continued until the time for the maximum intensity using various concentrations of yeast and dyes. The relaxation time τ, which corresponds to ~63·2% of the maximum fluorescence intensity, was shown to decrease to below 1 s with increasing DAPI concentration, whereas it remained constant for 2–3 s with increasing Auramine‐O concentration, for example at a yeast concentration of 100 µg ml−1. Conclusions For the conditions of yeast >10 µg ml−1, DAPI >1 µg ml−1 and Auramine‐O >0·1 µg ml−1, τ could be adjusted to below 5 s to achieve a rapid and stable staining. Significance and Impact of the Study Design and operating conditions for rapid and online measurements of microbes can be optimized.

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Section: Discussionsupporting

confidence: 87%

Section: Methodsmentioning

confidence: 99%

Optimal fluorescent‐dye staining time for the real‐time detection of microbes: a study of Saccharomyces cerevisiae

et al. 2020

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“…2c ). Another small-molecule dye, Auramine O (AO), was recently reported to generate strong fluorescence when mixed with DAP-10 30 and notably was used to study mycobacteria-induced neutrophil phagocytosis in human plasma 31 . In our investigations, fluorescence enhancement of AO without DAP-10 was minor in biological media (2 to 10-fold), whereas that with DAP-10 was significant in both FAB (3,800-fold) and biological media (82 to 494-fold, Supplementary Fig.…”

Section: Resultsmentioning

confidence: 99%

MicroRNA Detection in Biological Media Using a Split Aptamer Platform

Wang

Hast

Aggarwal

et al. 2022

Preprint

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Intercellular microRNA (miRNA)-based communication has been implicated in a wide array of functional and dysfunctional biological processes. This has raised attention to the potential use of miRNAs as biomarkers for disease diagnosis and prognosis and produced interest in their detection. Though the list of clinically significant miRNA biomarkers is rapidly expanding, it remains challenging to adapt current tools to investigate new targets in biological environments. Systematic approaches for the rapid development of miRNA biosensors are valuable to reduce this disparity. We describe here a methodology for developing aptamer-based fluorescent biosensors that can specifically detect miRNAs in biological environments, including culture medium from HeLa cells, human serum, and human plasma. This methodology includes the semi-rational design of the hybridization between a pair of split DNA aptamer oligonucleotides and the miRNA target to build a pool of potential sensor designs, and the screening of this pool for designs with high signal-to-background ratio and sequence selectivity. The method uses natural oligonucleotides without chemical modification, and is effective in buffer, 10%, and 30% (v/v) biological media. Following this approach, we developed sensors that detect three miRNA targets (miR-19b, miR-21, and miR-92a) at concentrations as low as 5 nM without amplification and are selective against single-nucleotide mutants. This work expands upon the current design principles of nucleic acid-based biosensors and provides a method to rapidly develop diagnostic tools for novel and niche miRNA targets of interest.

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“…Further enhancement in emission intensity of this red‐shifted band is observed with increasing PSS concentration, reaching saturation at 0.2 μM of PSS with an overall enhancement factor of ∼100. To this end, emission enhancement for AuO has been reported in various media like viscous solvents, confined environments, and recently, in the presence of some essential bioanalytes, such as, Insulin amyloid fibrils, G‐quadraplexes, etc [55–57] . The fluorescence enhancement of AuO in the presence of these analytes has been assigned to the suppression of intermolecular torsional relaxation in its excited state, compared to a feeble emission yield attained for AuO in its free state in low viscous solvents or in aqueous solutions [58,59] .…”

Section: Resultsmentioning

confidence: 99%

Supramolecular Control on the Optical Properties of a Dye‐Polyelectrolyte Assembly

2021

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Control of fluorescent molecular assemblies is an exciting area of research with large potential for various important applications, such as, fluorescence sensing/probing, cell imaging and monitoring drug‐delivery. In the present contribution, we have demonstrated control on the extent of aggregation of a dye‐polyelectrolyte assembly using a macrocyclic host molecule, sulfobutylether‐β‐cyclodextrin (SBE‐β‐CD). Initially, a cationic molecular rotor based organic dye, Auramine‐O (AuO), undergoes aggregation in the presence of an anionic polyelectrolyte, polystyrene sulfonate (PSS), and displays a broad intense new emission band along with large variation in its absorption features and excited‐state lifetime. A manipulation of the monomer‐aggregate equilibrium of the dye‐polyelectrolyte assembly has been achieved by introducing a cyclodextrin based supramolecular host, SBE‐β‐CD, which leads to relocation of AuO molecules from polyelectrolyte (PSS) to supramolecular host cavity, owing to the formation of a host‐guest complex between AuO and SBE‐β‐CD. A reversible control on this manipulation of monomer‐aggregate equilibrium is further achieved by introducing a competitive guest for the host cavity i. e., 1‐Adamantanol. Thus, we have demonstrated an interesting control on the dye‐polyelectrolyte aggregate assembly using a supramolecular host molecule which open up exciting possibilities to construct responsive materials using a repertoire of various host‐specific guest molecules.

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A label-free fluorescent molecular switch for a DNA hybridization assay utilizing a G-quadruplex-selective auramine O

Cited by 26 publications

References 33 publications

Optimal fluorescent‐dye staining time for the real‐time detection of microbes: a study of Saccharomyces cerevisiae

Optimal fluorescent‐dye staining time for the real‐time detection of microbes: a study of Saccharomyces cerevisiae

MicroRNA Detection in Biological Media Using a Split Aptamer Platform

Supramolecular Control on the Optical Properties of a Dye‐Polyelectrolyte Assembly

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