CRISPR/Cas-powered nanobiosensors for diagnostics

Biosensors and Bioelectronics, Год журнала: 2021, Номер 197, С. 113732 - 113732

Опубликована: Окт. 29, 2021

Язык: Английский

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Metal Ion-Regulated Fluorescent Sensor Array Based on Gold Nanoclusters for Physiological Phosphate Sensing

Analytical Chemistry, Год журнала: 2024, Номер 96(10), С. 4224 - 4231

Опубликована: Фев. 29, 2024

The detection of physiological phosphates (PPs) is great importance due to their essential roles in numerous biological processes, but the efficient different PPs simultaneously remains challenging. In this work, we propose a fluorescence sensor array for detecting based on metal-ion-regulated gold nanoclusters (AuNCs) via an indicator-displacement assay. Zn2+ and Eu3+ are selected assemble with two AuNCs, resulting quenching or enhancing fluorescence. Based competitive interaction metal ions AuNCs PPs, will be recovered owing disassembly AuNC-metal ion ensembles. Depending PPs' distinct responses, four-channel was established. not only exhibits good discrimination capability eight kinds (i.e., ATP, ADP, AMP, GTP, CTP, UTP, PPi, Pi) linear discriminant analysis also enables quantitative single phosphate (e.g., ATP) presence interfering mixtures. Moreover, potential application present real samples cell lysates serum) successfully demonstrated performance. This work illustrates ion-regulated as new sensing platform differential well other disease-related biomolecules.

Язык: Английский

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Holographic Optical Tweezers and Boosting Upconversion Luminescent Resonance Energy Transfer Combined Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas12a Biosensors

ACS Nano, Год журнала: 2021, Номер 15(5), С. 8142 - 8154

Опубликована: Янв. 11, 2021

Taking advantage of outstanding precision in target recognition and trans-cleavage ability, the recently discovered CRISPR/Cas12a system provides an alternative opportunity for designing fluorescence biosensors. To fully exploit analytical potential, we introduce here some meaningful concepts. First, collateral cleavage is efficiently activated a functional DNA regulation manner bottleneck which largely applicable to nucleic acids detection broken. After selection representative aptamer DNAzyme as transduction pathways, sensing coverage extended small organic compound (ATP) metal ion (Na+). The assay sensitivity significantly improved by utilizing bead-supported enrichment strategy wherein emerging holographic optical tweezers are used enhance imaging stability simultaneously achieve multiflux analysis. Last, sandwich-structured energy-concentrating upconversion nanoparticle triggered boosting luminescent resonance energy transfer mode comined face with complicated biological samples skillfully confining emitters into very limited inner shell. Following above attempts, developed biosensors not only present ultrasensitive behavior toward these model non-nucleic acid analytes but also can serve formidable toolbox determining real including single cell lysates human plasma, proving good practical application capacity.

Язык: Английский

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An Integrated Electrochemical Nanodevice for Intracellular RNA Collection and Detection in Single Living Cell

Angewandte Chemie International Edition, Год журнала: 2020, Номер 60(24), С. 13244 - 13250

Опубликована: Дек. 19, 2020

New tools for single-cell interrogation enable deeper understanding of cellular heterogeneity and associated behaviors functions. Information RNA expression in single cell could contribute to our knowledge the genetic regulatory circuits molecular mechanism disease development. Although significant progresses have been made intracellular analysis, existing methods a trade-off between operational complexity practical feasibility. We address this challenge by combining ionic current rectification property nanopipette reactor with duplex-specific nuclease-assisted hybridization chain reaction signal amplification realize simple nanosensor minimal invasiveness, which enables collection electrochemical detection cell-context preservation. Systematic studies on differentiation oncogenic miR-10b levels non-malignant breast cells, metastatic cancer cells as well non-metastatic were then realized nanotool accompanied assessment different drugs effects. This work has unveiled ability electrochemistry probe opened new opportunities study gene heterogeneous under physiological conditions down level.

Язык: Английский

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A boosting upconversion luminescent resonance energy transfer and biomimetic periodic chip integrated CRISPR/Cas12a biosensor for functional DNA regulated transduction of non-nucleic acid targets

Biosensors and Bioelectronics, Год журнала: 2020, Номер 169, С. 112650 - 112650

Опубликована: Сен. 26, 2020

Язык: Английский

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Gold Nanocluster-Based Fluorescent Microneedle Platform toward Visual Detection of ATP

Analytical Chemistry, Год журнала: 2023, Номер 95(32), С. 12104 - 12112

Опубликована: Авг. 1, 2023

Adenosine triphosphate (ATP) participates in the regulation of most biological processes, and ATP level is closely associated with many diseases. However, it still remains challenging to achieve on-site monitoring an equipment-free efficient way. Microneedles, a minimally invasive technology that can extract biomarkers from liquid biopsies, have recently emerged as useful tools for early diagnosis broad range In this work, we developed hydrogel microneedles are loaded ATP-specific dual-emitting gold nanoclusters (RhE-AuNCs) fast sampling on-needle detection ATP. These RhE-AuNCs were photo-crosslinked matrix form fluorescent microneedle patch. Based on ATP-induced Förster resonance energy transfer RhE-AuNCs, highly selective, sensitive, reliable sensor was developed. Moreover, simultaneous capture visual achieved by AuNC-loaded sensing platform, which exhibits promising performance. This work provides new approach design point-of-care also holds great potential further development microneedle-based analytical devices.

Язык: Английский

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Glutathione Content Detection of Single Cells under Ingested Doxorubicin by Functionalized Glass Nanopores

Analytical Chemistry, Год журнала: 2021, Номер 93(9), С. 4240 - 4245

Опубликована: Фев. 24, 2021

Glutathione (GSH) level in cells maintains redox homeostasis, so the detection of intracellular GSH is crucial for many pathological processes. In present study, we developed a cystamine-functionalized glass nanopore (cG-nanopore) specific single through Michael addition reaction between amino group cystamine and polydopamine adhered to inner wall nanopore. The cellular will cleave disulfide bond induce negative charge increase on residual surface modified nanopore, thus producing sensitive response current–voltage curves. sensing platform showed good relationship with concentration an excellent selectivity against interfering substances cells. variation content HeLa H8 incubated DOX was then further quantified by this method.

Язык: Английский

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A perspective on astrocyte regulation of neural circuit function and animal behavior

Glia, Год журнала: 2022, Номер 70(8), С. 1554 - 1580

Опубликована: Март 17, 2022

Studies over the past two decades have demonstrated that astrocytes are tightly associated with neurons and play pivotal roles in neural circuit development, operation, adaptation health disease. Nevertheless, precisely how integrate diverse neuronal signals, modulate structure function at multiple temporal spatial scales, influence animal behavior or disease through aberrant excitation molecular output remains unclear. This Perspective discusses new state-of-the-art approaches, including fluorescence indicators, opto- chemogenetic actuators, genetic targeting tools, quantitative behavioral assays, computational methods, might help resolve these longstanding questions. It also addresses complicating factors interpreting astrocytes' role regulation behavior, such as their heterogeneity, metabolism, inter-glial communication. Research on questions should provide a deeper mechanistic understanding of astrocyte-neuron assemblies' function, complex behaviors,

Язык: Английский

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Glass Capillary-Based Nanopores for Single Molecule/Single Cell Detection

ACS Sensors, Год журнала: 2023, Номер 8(2), С. 427 - 442

Опубликована: Янв. 20, 2023

A glass capillary-based nanopore (G-nanopore), due to its tapered tip, easy tunability in orifice size, and especially flexible surface modifications that can be tailored effectively capture enhance the ionic current signal of single entities (single molecules, cells, particles), offers a powerful nanoconfined sensing platform for diverse biological measurements cells molecules. Compared with other artificial two-dimensional solid-state nanopores, conical tip high spatial temporal resolution characteristics facilitate noninvasive molecule selected area (subcellular) cell detections (e.g., DNA mutations, highly expressed proteins, small markers reflect change tumor), as G-nanopore (≤100 nm) does negligible damage functions membrane integrity when inserted through membrane. In this brief review, we summarize preparation G-nanopores discuss advantages them platforms detection applications well cancer diagnosis treatment applications. We also describe bottlenecks limit widespread use clinical provide an outlook on future developments. The review will reader quick survey field rapid development tumor personalized medicine based single-molecule/single-cell bioassay.

Язык: Английский

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Target-Triggered Assembly in a Nanopipette for Electrochemical Single-Cell Analysis

Analytical Chemistry, Год журнала: 2020, Номер 93(2), С. 1200 - 1208

Опубликована: Дек. 10, 2020

Engineered nanopipette tools have recently emerged as a powerful approach for electrochemical nanosensing, which has major implications in both fundamental biological research and biomedical applications. Herein, we describe generic method of target-triggered assembly aptamers is exemplified by sensitive rapid single-cell analysis adenosine triphosphate (ATP), ubiquitous energy source life important signaling molecules many physiological processes. Specifically, layer thiolated immobilized onto Au-coated interior wall tip. With backfilled pairing aptamers, the engineered then used probing intracellular ATP via ATP-dependent linkage split aptamers. Due to higher surface charge density from aptamer assembly, nanosensor would exhibit an enhanced rectification signal. Besides, this ATP-responsive tool possesses excellent selectivity stability well high recyclability. This work provides practical capable analysis. More generally, integrated with other recognition elements, proposed mechanism could serve viable basis addressing species.

Язык: Английский

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