Principal factors that determine the extension of detection range in molecular beacon aptamer/conjugated polyelectrolyte bioassays

Jeong, Ji Eun; Kim, Boram; Woo, Shinjae; Hwang, Sungu; Bazan, Guillermo C.; Woo, Han Young

doi:10.1039/c4sc03258f

Cited by 11 publications

(5 citation statements)

References 57 publications

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“…Fluorescence resonance energy transfer (FRET) can be utilized for multicolor imaging because a single donor excitation can result in fluorescence emission of various acceptor dyes. FRET involves nonradiative energy transfer between donor and acceptor molecules via dipole–dipole coupling and has been widely used for fluorescent and ratiometric sensing. − Because the FRET efficiency is inversely proportional to the donor–acceptor intermolecular distance raised to the sixth power, Pdots that encage the fluorophores within the nanosized particles can be an appropriate platform to locate donor and acceptor dyes in close proximity. Furthermore, the photoluminescence (PL) of red- and near-infrared (NIR)-emitting probes can be enhanced via the antenna effect of CPs, which can be advantageous for biological imaging owing to the deep tissue penetration and negligible autofluorescence.…”

Section: Introductionmentioning

confidence: 99%

Green-, Red-, and Near-Infrared-Emitting Polymer Dot Probes for Simultaneous Multicolor Cell Imaging with a Single Excitation Wavelength

et al. 2020

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We report newly synthesized fluorescence resonance energy transfer (FRET)-based green-, red-, and near-infrared (NIR)-emitting polymer dot (Pdot) probes. Fluorescent Pdots (∼60 nm) were prepared with a green-emissive conjugated polymer (PPDT-P, donor) alone or mixing the donor with a red-or NIRemitting fluorophore (T-DCS or ITIC, acceptor), where an optically inert matrix polymer [poly(styrene-co-maleic anhydride)] was mixed together to minimize the aggregation-caused quenching by diluting the fluorophores and surface functionalization for further bioconjugation with antibodies for active targeting. Highly fluorescent green-emissive PPDT-P Pdots were prepared with a photoluminescence (PL) quantum efficiency of ∼30% and the FRETmediated red and NIR PL was intensified by 5.3−8.5 times (with high FRET ratios of ∼9) via the efficient energy transfer (FRET efficiency of 80−98%) and antenna effect, compared with the signals obtained via direct excitation of the fluorophores in Pdots. All three types of Pdot/antibody conjugates were simultaneously immunostained to COS-7 cells (showing minimal cross-reactivity), reducing the tedious sequential immunostaining process to a single step. Finally, we obtained high-contrast three-color cell images with little spectral leakthrough by exciting all the probes simultaneously at a single wavelength at 405 nm without the need for a complicated or expensive multiple-excitation setup.

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Section: Introductionmentioning

confidence: 99%

Green-, Red-, and Near-Infrared-Emitting Polymer Dot Probes for Simultaneous Multicolor Cell Imaging with a Single Excitation Wavelength

et al. 2020

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“…In previous works, the adenosine-induced aptamer conformational switch and DNA duplex dissociation have been used for design of ATP-responsive sensor, the disassembly of nanoparticles, and controlled drug release. [48][49][50] Here, to investigate whether the surface property of hydrogel could be programmed with ATP molecules, we labelled the cDNA with a uorescent dye, uorescein isothiocyanate (FITC) (Fig. 2a).…”

Section: Resultsmentioning

confidence: 99%

“…In our strategy, hydrogel was chosen as the supporting substrate due to its excellent abilities to simulate natural extracellular matrix. The ATP aptamer (Apt-DNA) [48][49][50] and its complementary DNA (cDNA) were rst conjugated onto the hydrogel. They acted as intelligent mediators to immobilize the cell-adhesive RGD ligand.…”

Section: Introductionmentioning

confidence: 99%

Endogenous signalling control of cell adhesion by using aptamer functionalized biocompatible hydrogel

Wang

Ren

et al. 2015

Chem. Sci.

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“…A MBA containing the ATP‐specific aptamer sequence in its loop part (GGGG AGTA TTGC GGAG GA) with a fluorophore (6‐FAM) and quencher (DABCYL) in both termini was selected as the ATP probe (Figure ) . The ATP‐specific MBA probe preferentially takes a hairpin conformation via hydrogen bonding interactions in its stem part.…”

Section: Resultsmentioning

confidence: 99%

Modulation of Charge Density of Cationic Conjugated Polyelectrolytes for Improving the FRET‐Induced Sensory Signal with Enhanced On/Off Ratio

Jeong

Jung

Kim

et al. 2015

Macro Chemistry & Physics

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Three types of cationic polyfl uorenes with a different number of ionic groups are synthesized to modulate their solubility in water and the interactions with anionic oligonucleotide probes (DNA-C). Increasing the charge density improves the water-solubility of conjugated polyelectrolytes (CPEs) but decreases its optical antenna effect in the electrostatic sensory platform (CPE/DNA-C), resulting in low fl uorescence resonance energy transfer (FRET) signal. With increasing charge density in CPE, the concentration of conjugated units in CPE/DNA-C decreases at [+]:[−] = 1:1, after which little driving force is expected to bring positive CPEs and negative DNA-C into close proximity for effi cient FRET. CPF2 with 2 ionic groups (per repeat unit) and hydrophilic polyether sidechains exhibits good water-solubility and a higher FRET signal with an enhanced signal on/off ratio. In addition, the charge density in CPEs modulates the range of sensory responses by forming a tighter or weaker electrostatic CPE/DNA-C complex, which perturbs the probe-target binding interactions.

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Principal factors that determine the extension of detection range in molecular beacon aptamer/conjugated polyelectrolyte bioassays

Abstract: A new bioassay strategy based on the molecular beacon aptamer/conjugated polyelectrolyte demonstrates a fine-tuning of the detection range and limit of detection for weakly-binding targets.

Cited by 11 publications

References 57 publications

Green-, Red-, and Near-Infrared-Emitting Polymer Dot Probes for Simultaneous Multicolor Cell Imaging with a Single Excitation Wavelength

Green-, Red-, and Near-Infrared-Emitting Polymer Dot Probes for Simultaneous Multicolor Cell Imaging with a Single Excitation Wavelength

Endogenous signalling control of cell adhesion by using aptamer functionalized biocompatible hydrogel

Modulation of Charge Density of Cationic Conjugated Polyelectrolytes for Improving the FRET‐Induced Sensory Signal with Enhanced On/Off Ratio

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