An Enzymatically Gated Catalytic Hairpin Assembly Delivered by Lipid Nanoparticles for the Tumor‐Specific Activation of Signal Amplification in miRNA Imaging

Angewandte Chemie International Edition, Journal Year: 2022, Volume and Issue: 61(51)

Published: Nov. 16, 2022

MicroRNA (miRNA) imaging in disease sites is vital to elucidate their role cancer progression. However, limited tumor specificity remains a major barrier for traditional amplification approaches due associated background signal leakage. Here, we report generalizable approach via the combination of enzymatically triggered catalytic hairpin assembly with lipid nanoparticles (LNPs)-based delivery strategy tumor-specific activation and therefore sensitive miRNA imaging. The established engineering activated motifs achieve triggable cells. Furthermore, by introduction LNPs combat biological barriers, demonstrate that system enables amplified vivo reduced off-tumor signal, leading enhanced tumor-to-background contrast compared methods. This relies on specific triggers controlled distinguish cells from normal should be useful diagnosis.

Language: Английский

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Enzyme-Free Dynamic DNA Reaction Networks for On-Demand Bioanalysis and Bioimaging

Accounts of Chemical Research, Journal Year: 2024, Volume and Issue: unknown

Published: Jan. 25, 2024

ConspectusThe pursuit of in-depth studying the nature and law life activity has been dominating current research fields, ranging from fundamental biological studies to applications that concern synthetic biology, bioanalysis, clinical diagnosis. Motivated by this intention, spatiotemporally controlled in situ analysis living cells a prospective branch virtue high-sensitivity imaging key biomolecules, such as biomarkers. The past decades have attested deoxyribonucleic acid (DNA), with biocompatibility, programmability, customizable features, is competitive biomaterial for constructing high-performance molecular sensing tools. To conquer complexity wide extracellular–intracellular distribution biomarkers, it meaningful breakthrough explore high-efficiently amplified DNA circuits, which excel at operating complex yet captivating dynamic reaction networks various bioapplications. In parallel, multidimensional performance improvements nucleic including availability, detection sensitivity, reliability, are critical parameters realizing accurate cell regulation bioanalysis.In Account, we summarize our recent work on enzyme-free bioanalysis three main aspects: circuitry functional extension recognition epigenetic regulation, amplification ability improvement sensitive biomarker detection, site-specific activation systems reliable imaging. first part, designed an epigenetically responsive deoxyribozyme (DNAzyme) system intracellular gene enriches possible analyzed species chemically modifying conventional DNAzyme. For example, exquisite N6-methyladenine (m6A)-caged DNAzyme was built achieving precise FTO (fat mass obesity-associated protein)-directed regulation. addition, varieties DNAzyme-based nanoplatforms self-sufficient cofactor suppliers were assembled, subdued speed-limiting hardness cofactors live-cell applications. second developed series hierarchically assembled improve signal transduction traditional circuits. First, circuit significantly enhanced via several heterogeneously or homogeneously concatenated models. Furthermore, feedback pathway integrated into these thus dramatically increasing efficiency. Second, considering cellular environment, simplified redundancy multicomponents procedures cascaded relying minimal component merely one modular catalytic reaction, guaranteed high cell-delivering uniformity while fostering kinetics reliability. third constructed in-cell-selective endogenous-stimulated multiply recognitions, could not only eliminate leakage, but also retain its on-site multiplex amplification. Based strategy, more availability scenarios acquired These demonstrate purpose-to-concreteness engineering tailored multimolecule multiple amplification, high-gain high-reliability targeted bioanalysis. We envision network can contribute bioanalytical layouts, will facilitate progression diagnosis prognosis.

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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An Intelligent Redox‐Responsive DNA Circuit for Robust On‐Site Profiling of Glutathione‐MicroRNA Signaling Pathway

Advanced Functional Materials, Journal Year: 2024, Volume and Issue: 34(26)

Published: Feb. 22, 2024

Abstract Synthetic biochemical circuits (e.g., DNA circuits) remain at the forefront of intracellular biosensing tasks yet are hindered by undesired off‐site activation and accompanying signal leakage. Herein, study attempts to overcome this limitation developing a simple‐yet‐powerful endogenous glutathione (GSH)‐regulating tactic that permits robust distinguishable on‐site microRNA (miRNA) imaging under disturbed redox homeostasis. Specifically, hierarchically activated catalytic (HAD) circuit is fabricated grafting disulfide linkage within entropy‐driven circuitry (EDC) reactants. It exemplified HAD system promises spatiotemporally selective microRNA‐21 (miR‐21) in living cells differentiation tumor from normal cells. The correlationship between GSH miRNA extensively explored live cells, can substantially expand toolbox for profiling processes.

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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Tailoring a Minimal Self‐Replicate DNA Circuit for Highly Efficient Intracellular Imaging of microRNA

Small, Journal Year: 2023, Volume and Issue: 19(17)

Published: Jan. 30, 2023

Abstract Trace analyte detection in complex intracellular environment requires the development of simple yet robust self‐sufficient molecular circuits with high signal‐gain and anti‐interference features. Herein, a minimal non‐enzymatic self‐replicate DNA circuitry (SDC) system is proposed high‐signal‐gain for highly efficient biosensing living cells. It facilely engineered through self‐stacking only one elementary cascade hybridization reaction (CHR), thus encoding more economic effective amplification pathways reactants. Trigger ( T ) stimulates activation CHR producing numerous replica that reversely motivate new cycles, achieving successive self‐replication an exponentially magnified readout signal. The intrinsic circuity design self‐accelerated format SDC experimentally demonstrated theoretically simulated. With configuration low reactant complexity, amplifier enables high‐contrast accurate visualization microRNA (miRNA), ascribing to its recognition signal amplification, offering promising strategy monitoring these clinically significant analytes.

Language: Английский

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Stimuli‐Responsive DNA Circuits for High‐Performance Bioimaging Application

Advanced Sensor Research, Journal Year: 2023, Volume and Issue: 2(9)

Published: March 3, 2023

Abstract Probing endogenous molecules in living entities is significant to help decipher biological functions and exploit novel theranostics. DNA circuits that can recognize molecular inputs of interest transduce them into readable signal outputs an isothermal autonomous manner have been actively pursued as versatile toolkits for intracellular biosensing research. Tremendous efforts are being devoted developing integrated with high sensitivity, while spatiotemporal selectivity often overlooked the construction functional circuitry systems. This requires development stimuli‐responsive be activated on‐demand from initial sensing‐blunt state sensing‐ready under a programmable manner, achieving precise bioimaging control. In this review, overview recent advances respond particular triggers, including external physical stimuli cues, their spatiotemporally controllable applications provided. The current challenges potential solutions these future developments emerging field also discussed.

Language: Английский

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Endogenous AND Logic DNA Nanomachine for Highly Specific Cancer Cell Imaging

Analytical Chemistry, Journal Year: 2024, Volume and Issue: 96(18), P. 7030 - 7037

Published: April 24, 2024

Intracellular cancer-related biomarker imaging strategy has been used for specific identification of cancer cells, which was great importance to accurate clinical diagnosis and prognosis studies. Localized DNA circuits with improved sensitivity showed potential intracellular biomarkers imaging. However, the ability localized specifically image cells is limited by off-site signal leakage associated a single-biomarker sensing strategy. Herein, we integrated endogenous enzyme-powered logic-responsive amplifying capability construct self-assembled endogenously AND logic nanomachine (EDN) highly cell When EDN encountered cell, overexpressed repairing enzyme apurinic/apyrimidinic endonuclease 1 (APE1) miR-21 could synergistically activate circuit via cascaded toehold-mediated strand displacement (TMSD) reactions, resulting in amplified fluorescence resonance energy transfer (FRET) signal. In this strategy, both APE1 miR-21, served as two "keys" operation reduce off-tumor leakage. Such multiplied molecular recognition/activation powerful toolbox realized capture reliable biomolecules living cells.

Language: Английский

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A Cellular Membrane‐Confined Concatenate DNA Circuit for Non‐Invasive Cell Modulation with High Accuracy and Efficiency

Advanced Functional Materials, Journal Year: 2023, Volume and Issue: 33(40)

Published: June 15, 2023

Abstract Non‐invasive cell regulation represents a promising approach for on‐site modulation, but is confronted with inaccuracy or poor selectivity. Herein, by virtue of the interfacial‐activated concatenate DNA circuit, an activated recombination‐to‐modulation (ARM) machinery developed to achieve accurate membrane imaging and effective modulation. Bioorthogonal signal transduction through catalytic hairpin assembly circuit can continuously regenerate triggers activating subsequent no activator consumption occupation. The ARM strategy utilizes amplified hybridization chain reaction sensitively rearrange receptors blocking related signaling pathways indirectly modulating behaviors. Based on these two receptor inputs, controllably‐activated enables identification target cells among similar interfering high accuracy sensitivity. This has demonstrated good performance in efficient inhibition tumor metastasis via disruption HGF/c‐Met‐signaling pathway. modular‐designed provides general platform non‐invasive specific suggesting broad opportunity advanced exploration cellular metabolism

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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