Interactions of Potential Protein Cancer Biomarker Survivin with Plasmonic Nanoparticles and Its Dynamics in Cancer Cells Studied Using Fluorescence Molecular-Beacon Probes, Gated-RET and EQCN Methods

2015 ◽  
Vol 1720 ◽  
Author(s):  
Magdalena Stobiecka ◽  
Agata Chalupa ◽  
Beata Dworakowska
Keyword(s):  
Molecular Beacon ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Fluorescein Isothiocyanate ◽  
Cancer Biomarker ◽  
Molecular Beacons ◽  
Target Sequence ◽  
Mrna Levels ◽  
Loop Area ◽  
Fluorescence Turn On

ABSTRACTThe protein survivin (Sur) has been considered as a potential cancer biomarker due to its involvement in disrupting normal cell cycle by stimulating proliferation and inhibiting cell apoptosis. In this work, we have focused on exploring novel platforms for sensitive monitoring of Sur expression, based on molecular beacons and protein modulation of plasmon-controlled fluorescence. In this framework, we show that Sur can be employed in gating the resonance energy transfer (gRET) between fluorescein isothiocyanate probe dye (FITC) and plasmonic citrate-capped gold nanoparticles (AuNP@Cit). Furthermore, we have designed fluorescent dye-bearing molecular beacons (MBs) targeting nucleotides of the survivin mRNA. The antisense oligonucleotide complementary to the target sequence, inserted in the loop area of the hairpin MB structure, has enabled the fluorescence turn-ON MB switching in the presence of the target, thus signaling the high Sur mRNA levels and enhanced Sur protein expression.

scholarly journals CRISPR/dual-FRET molecular beacon for sensitive live-cell imaging of non-repetitive genomic loci

2019 ◽  
Vol 47 (20) ◽  
pp. e131-e131 ◽  
Author(s):  
Shiqi Mao ◽  
Yachen Ying ◽  
Xiaotian Wu ◽  
Christopher J Krueger ◽  
Antony K Chen
Keyword(s):  
Live Cell Imaging ◽  
Cell Imaging ◽  
Molecular Beacon ◽  
Fluorescent Protein ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Dynamic Imaging ◽  
Molecular Beacons ◽  
Imaging Systems ◽  
Guide Rna

Abstract Clustered regularly interspaced short palindromic repeats (CRISPR)-based genomic imaging systems predominantly rely on fluorescent protein reporters, which lack the optical properties essential for sensitive dynamic imaging. Here, we modified the CRISPR single-guide RNA (sgRNA) to carry two distinct molecular beacons (MBs) that can undergo fluorescence resonance energy transfer (FRET) and demonstrated that the resulting system, CRISPR/dual-FRET MB, enables dynamic imaging of non-repetitive genomic loci with only three unique sgRNAs.

A multiplex fluorophore molecular beacon: detection of the target sequence using large Stokes shift and multiple emission signal properties

2015 ◽  
Vol 51 (14) ◽  
pp. 2939-2942 ◽  
Author(s):  
Han Na Joo ◽  
Young Jun Seo
Keyword(s):  
Energy Transfer ◽  
Fluorescence Resonance Energy Transfer ◽  
Molecular Beacon ◽  
Stokes Shift ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Fluorescence Properties ◽  
Target Sequence ◽  
Large Stokes Shift ◽  
Emission Signal

We have developed a multiplex fluorophore molecular beacon (mfMB) with fluorophores located at its end to produce unique FRET (Fluorescence Resonance Energy Transfer). It exhibited diverse fluorescence properties depending on the mixing pattern, such as large Stokes shift emission and multiple colors.

Spectroscopic Features of Dual Fluorescence/Luminescence Resonance Energy-Transfer Molecular Beacons

2003 ◽  
Vol 75 (15) ◽  
pp. 3697-3703 ◽  
Author(s):  
Andrew Tsourkas ◽  
Mark A. Behlke ◽  
Yangqing Xu ◽  
Gang Bao
Keyword(s):  
Energy Transfer ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Molecular Beacons ◽  
Dual Fluorescence

scholarly journals Ratiometric pH Sensing and Imaging in Living Cells with Dual-Emission Semiconductor Polymer Dots

Molecules ◽  
2019 ◽  
Vol 24 (16) ◽  
pp. 2923
Author(s):  
Piaopiao Chen ◽  
Iqra Ilyas ◽  
Su He ◽  
Yichen Xing ◽  
Zhigang Jin ◽  
...  
Keyword(s):  
Fluorescent Dyes ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Fluorescein Isothiocyanate ◽  
Water Soluble ◽  
Ph Sensing ◽  
Dual Emission ◽  
Polymer Dots ◽  
Ratiometric Ph Sensing

Polymer dots (Pdots) represent newly developed semiconductor polymer nanoparticles and exhibit excellent characteristics as fluorescent probes. To improve the sensitivity and biocompatibility of Pdots ratiometric pH biosensors, we synthesized 3 types of water-soluble Pdots: Pdots-PF, Pdots-PP, and Pdots-PPF by different combinations of fluorescent dyes poly(9,9-dioctylfluorenyl-2,7-diyl) (PFO), poly[(9,9-dioctyl-fluorenyl-2,7-diyl)-co-(1,4-benzo-{2,1′,3}-thiadazole)] (PFBT), and fluorescein isothiocyanate (FITC). We found that Pdots-PPF exhibits optimal performance on pH sensing. PFO and FITC in Pdots-PPF produce pH-insensitive (λ = 439 nm) and pH-sensitive (λ = 517 nm) fluorescence respectively upon a single excitation at 380 nm wavelength, which enables Pdots-PPF ratiometric pH sensing ability. Förster resonance energy transfer (FRET) together with the use of PFBT amplify the FITC signal, which enables Pdots-PPF robust sensitivity to pH. The emission intensity ratio (I517/I439) of Pdots-PPF changes linearly as a function of pH within the range of pH 3.0 to 8.0. Pdots-PPF also possesses desirable reversibility and stability in pH measurement. More importantly, Pdots-PPF was successfully used for cell imaging in Hela cells, exhibiting effective cellular uptake and low cytotoxicity. Our study suggests the promising potential of Pdots-PPF as an in vivo biomarker.

A paper-supported sandwich immunosensor based on upconversion luminescence resonance energy transfer for the visual and quantitative determination of a cancer biomarker in human serum

The Analyst ◽  
2020 ◽  
Vol 145 (12) ◽  
pp. 4181-4187 ◽  
Author(s):  
Mengyuan He ◽  
Ning Shang ◽  
Lin Shen ◽  
Zhihong Liu
Keyword(s):  
Energy Transfer ◽  
Quantitative Determination ◽  
Human Serum ◽  
Upconversion Luminescence ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Cancer Biomarker

A portable and sensitive paper-supported sandwich immunosensor based on UC-FRET was developed for the visual and quantitative determination of CEA.

scholarly journals Molecular beacons can assess changes in expression and 3′-polyadenylation of human eNOS mRNA

2009 ◽  
Vol 296 (3) ◽  
pp. C498-C504 ◽  
Author(s):  
Rachel Jones ◽  
Meredith B. Baker ◽  
Martina Weber ◽  
David G. Harrison ◽  
Gang Bao ◽  
...  
Keyword(s):  
Gene Expression ◽  
Intracellular Signaling ◽  
Molecular Beacon ◽  
Competitive Inhibition ◽  
Molecular Beacons ◽  
Mrna Levels ◽  
Vascular Homeostasis ◽  
Enos Mrna ◽  
Endothelial Gene Expression ◽  
Hybridization Assays

The endothelium plays an essential role in maintaining vascular homeostasis, and it fulfills this role by modulating intracellular signaling and gene expression in response to chemical and mechanical stimuli. Assessing changes in endothelial gene expression is essential to understanding how physiological and pathophysiological processes modulate vascular homeostasis. Here we describe the use of molecular beacons to rapidly and quantitatively assess expression and 3′-polyadenylation of a gene that is important for vascular homeostasis, endothelial nitric oxide synthase (eNOS). Single- and dual-fluorescence resonance energy transfer (FRET) molecular beacon hybridization assays were developed to measure changes in mRNA levels and 3′-polyadenylation, respectively, in primary human endothelial cell cultures subjected to laminar shear stress or statin treatment. Optimized beacon hybridization assays took ∼15 min to perform, and eNOS mRNA levels were validated by quantitative real-time RT-PCR. Competitive inhibition assays and posttranscriptional silencing of eNOS expression were used to verify the specificity of molecular beacon fluorescence. Finally, the dual-FRET method was used to assess eNOS polyadenylation in tissues isolated from mice subjected to exercise training. These data demonstrate that molecular beacons can be used to rapidly and efficiently measure endothelial gene expression and 3′-polyadenylation. This approach could easily be adapted for studies of other endothelial genes and has promise for applications in live endothelial cells.

Sign in / Sign up

Export Citation Format

Share Document