An electrochemical biosensor designed with entropy-driven autocatalytic DNA circuits for sensitive detection of ovarian cancer-derived exosomes

Biosensors and Bioelectronics, Journal Year: 2024, Volume and Issue: 250, P. 116060 - 116060

Published: Jan. 23, 2024

Language: Английский

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Photoactivatable Engineering of CRISPR/Cas9‐Inducible DNAzyme Probe for In Situ Imaging of Nuclear Zinc Ions

Angewandte Chemie International Edition, Journal Year: 2024, Volume and Issue: 63(10)

Published: Jan. 23, 2024

DNAzyme-based fluorescent probes for imaging metal ions in living cells have received much attention recently. However, employing situ within subcellular organelles, such as nucleus, remains a significant challenge. We developed three-stranded DNAzyme probe (TSDP) that contained 20-base-pair (20-bp) recognition site of CRISPR/Cas9, which blocks the activity. When Cas9, with its specialized nuclear localization function, forms an active complex sgRNA cell it cleaves TSDP at site, resulting formation catalytic structure. With this design, CRISPR/Cas9-inducible Zn

Language: Английский

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Spatially Localized Entropy-Driven Evolution of Nucleic Acid-Based Constitutional Dynamic Networks for Intracellular Imaging and Spatiotemporal Programmable Gene Therapy

Journal of the American Chemical Society, Journal Year: 2024, Volume and Issue: 146(30), P. 20685 - 20699

Published: July 16, 2024

The primer-guided entropy-driven high-throughput evolution of the DNA-based constitutional dynamic network, CDN, is introduced. entropy gain associated with process provides a catalytic principle for amplified emergence CDN. concept applied to develop programmable, spatially localized DNA circuit effective in vitro and vivo theranostic, gene-regulated treatment cancer cells. consists tetrahedron core modified at its corners four tethers that include encoded base sequences exhibiting capacity emerge assemble into [2 × 2] Two are caged by pair siRNA subunits, blocking mute, dynamically inactive configuration. In presence miRNA-21 as primer, subunits displaced, resulting release siRNAs silencing HIF-1α mRNA fast reconfiguration CDN is, however, engineered be reconfigured miRNA-155 an equilibrated mixture enriched DNAzyme component, catalyzing cleavage EGR-1 mRNA. nanostructure stimulates enhanced permeation miRNA-triggered leads cooperative bis-gene-silencing mRNAs, selective apoptosis breast cells inhibition tumors tumor bearing mice.

Language: Английский

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Scaling up of a Self‐Confined Catalytic Hybridization Circuit for Robust microRNA Imaging

Advanced Science, Journal Year: 2024, Volume and Issue: 11(22)

Published: April 13, 2024

Abstract The precise regulation of cellular behaviors within a confined, crowded intracellular environment is highly amenable in diagnostics and therapeutics. While synthetic circuitry system through concatenated chemical reaction network has rarely been reported to mimic dynamic self‐assembly system. Herein, catalytic self‐defined circuit (CSC) for the hierarchically assembly DNA domino nanostructures engineered. By incorporating pre‐sealed symmetrical fragments into preying hairpin reactants, CSC allows hierarchical via microRNA (miRNA)‐powered self‐sorting hybridization reaction. With minimal strand complexity, this self‐sustainable streamlined component achieved localization‐intensified cascaded signal amplification. Profiting from self‐adaptively reaction, reliable robust method discriminating carcinoma tissues corresponding para‐carcinoma tissues. CSC‐sustained strategy provides comprehensive smart toolbox organizing various nanostructures, which may facilitate more insights clinical diagnosis therapeutic assessment.

Language: Английский

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Multiply Guaranteed Catalytic DNA Circuit for Cancer-Cell-Selective Imaging of miRNA and Robust Evaluation of Drug Resistance

Analytical Chemistry, Journal Year: 2024, Volume and Issue: 96(14), P. 5560 - 5569

Published: March 26, 2024

Catalytic DNA circuits are desirable for sensitive bioimaging in living cells; yet, it remains a challenge to monitor these intricate signal communications because of the uncontrolled circuitry leakage and insufficient cell selectivity. Herein, simple yet powerful DNA-repairing enzyme (APE1) activation strategy is introduced achieve site-specific exposure catalytic circuit realizing selectively amplified imaging intracellular microRNA robust evaluation APE1-involved drug resistance. Specifically, reactants firmly blocked by recognition/cleavage site prevent undesirable off-site leakage. The caged has no target-sensing activity until its components activated via enzyme-mediated structural reconstitution finally transduces fluorescence within miRNA stimulation. designed demonstrates an enhanced signal-to-background ratio assay as compared with conventional enables cancer-cell-selective miRNA. In addition, shows sensing performance visualizing APE1-mediated chemoresistance cells, which anticipated in-depth clinical diagnosis chemotherapy research.

Language: Английский

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Toehold Strand Displacement-Mediated Exponential HCR for Highly Sensitive and Specific Analysis of miRNA in Living Cells

Analytical Chemistry, Journal Year: 2024, Volume and Issue: 96(22), P. 9078 - 9087

Published: May 21, 2024

As an important disease biomarker, the development of sensitive detection strategies for miRNA, especially intracellular miRNA imaging strategies, is helpful early diagnosis diseases, pathological research, and drug development. Hybridization chain reaction (HCR) widely used analysis because its high specificity lack biological enzymes. However, classic HCR exhibits linear amplification with low efficiency, limiting use rapid trace in living cells. To address this problem, we proposed a toehold-mediated exponential (TEHCR) to achieve highly efficient cells using β-FeOOH nanoparticles as transfection vectors. The limit TEHCR was 92.7 fM, which 8.8 × 103 times lower compared traditional HCR, it can effectively distinguish single-base mismatch specificity. also different expression levels cancer normal Furthermore, be construct OR logic gates dual without need additional probes, demonstrating flexibility. This method expected play role clinical miRNA-related well promote gates.

Language: Английский

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A Methylation-Gated DNAzyme Circuit for Spatially Controlled Imaging of MicroRNA in Cells and Animals

Analytical Chemistry, Journal Year: 2024, Volume and Issue: 96(23), P. 9666 - 9675

Published: May 30, 2024

Epigenetic modification plays an indispensable role in regulating routine molecular signaling pathways, yet it is rarely used to modulate self-assembly networks. Herein, we constructed a bioorthogonal demethylase-stimulated DNA circuitry (DSC) system for high-fidelity imaging of microRNA (miRNA) live cells and mice by eliminating undesired off-site signal leakage. The simple robust DSC composed primary cell-specific regulation (CR) module ultimate signal-transducing amplifier (SA) module. After the modularly designed was delivered into target cells, DNAzyme CR site-specifically activated endogenous demethylase produce fuel strands subsequent miRNA-targeting SA Through on-site multiply guaranteed recognitions, lucid efficient realized reliably amplified vivo miRNA sensing enabled in-depth exploration demethylase-involved pathway with cells. Our bioorthogonally on-site-activated represents universal versatile biomolecular platform via various regulations shows more prospects different personalized theragnostics.

Language: Английский

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Dual miRNA-Triggered DNA Walker Assisted by APE1 for Specific Recognition of Tumor Cells

Analytical Chemistry, Journal Year: 2024, Volume and Issue: 96(17), P. 6774 - 6783

Published: April 18, 2024

The identification of a specific tumor cell is crucial for the early diagnosis and treatment cancer. However, it remains challenge due to limited sensitivity accuracy, long response time, low contrast recent approaches. In this study, we develop dual miRNA-triggered DNA walker (DMTDW) assisted by APE1 recognition cells. miR-10b miR-155 were selected as research models. Without presence, inactive its walking strand W locked L1 L2. After are input, triggered bind L2 W-L1-L2, respectively, unlocking W. driven endogenous that highly catalytic expressed in cytoplasm cells but barely normal cells, ensuring high reaction efficiency Dual miRNA input required trigger walker, making strategy with accuracy. DMTDW exhibited analysis detection limit 44.05 pM. Living cell-imaging experiments confirmed could effectively respond fluctuation specifically identified MDA-MB-231 from different lines. proposed sensitive, rapid, accurate recognition. We believe can become powerful diagnostic tool

Language: Английский

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Spatially Selective MicroRNA Imaging in Human Colorectal Cancer Tissues Using a Multivariate-Gated Signal Amplification Nanosensor

Journal of the American Chemical Society, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 11, 2025

MicroRNA (miRNA) is involved in the genesis viand development of colorectal cancer. The vivo imaging miRNA at tumor sites essential for understanding its role cancer pathology and therapeutic target identification. However, achieving accurate hindered by low abundance miRNAs cells nonspecific signal leakage normal tissues. Here, we report a multivariate-gated catalytic hairpin assembly (CHA) nanosensor specific amplified microRNA-21 (miR-21) human tissues to reveal underlying miR-21-associated molecular mechanism. endogenous glutathione exogenous near-infrared design combination with CHA probes improves strength miR-21 reduces background interference. enables vivo, signal-to-background ratios are 1.6-fold compared traditional methods. With assistance designed nanosensor, achieve preliminary identification from clinical surgical resection samples. overexpressed found suppress core mismatch repair recognition protein mutS homologue 2 DNA damage inhibit efficacy strategy probe design, which combines activation methods amplification system, applicable disease-relevant mechanism research.

Language: Английский

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Spatiotemporally Programmed Disassembly of Multifunctional Integrated DNAzyme Nanoplatfrom for Amplified Intracellular MicroRNA Imaging

Small, Journal Year: 2023, Volume and Issue: 20(2)

Published: Sept. 5, 2023

Abstract The sensing performance of DNAzymes in live cells is tremendously hampered by the inefficient and inhomogeneous delivery DNAzyme probes their incontrollable off‐site activation, originating from susceptibility to nuclease digestion. This requires development a more compact robust DNAzyme‐delivering system with site‐specific activation property. Herein, highly Zn@DDz nanoplatform constructed integrating unimolecular microRNA‐responsive DNA‐cleaving (DDz) probe requisite Zn 2+ ‐ion cofactors, amplified intracellular imaging microRNA via spatiotemporally programmed disassembly nanoparticles achieved. multifunctional simply composed structurally blocked self‐hydrolysis DDz inorganic bridge, high loading capacity, can effectively deliver initially catalytic inert into living enhanced stabilities. Upon entry acidic microenvironment cells, self‐sufficient nanoparticle disassembled release simultaneously supply cofactors. Then, endogenous microRNA‐21 catalyzes reconfiguration for generating readout signal multiply guaranteed performance. Thus, this work paves an effective way promoting DNAzyme‐based biosensing systems shows great promise clinical diagnosis.

Language: Английский

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