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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Modular Weaving DNAzyme in Skeleton of DNA Nanocages for Photoactivatable Catalytic Activity Regulation

Angewandte Chemie International Edition, Journal Year: 2024, Volume and Issue: 63(18)

Published: March 22, 2024

Abstract DNAzymes exhibit tremendous application potentials in the field of biosensing and gene regulation due to its unique catalytic function. However, spatiotemporally controlled DNAzyme activity remains a daunting challenge, which may cause nonspecific signal leakage or silencing systems. Here, we report photochemical approach via modular weaving active into skeleton tetrahedral DNA nanocages (TDN) for light‐triggered on‐demand liberation thus conditional control activity. We demonstrate that direct encoding TDN could improve biostability ensure delivery efficiency, comparing with conventional surface anchoring strategy. Furthermore, molecular nanostructures allows remote DNAzyme‐mediated high spatiotemporal precision light. In addition, is applicable editing functions other functional nucleic acids.

Language: Английский

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DNA Nanotechnology Targeting Mitochondria: From Subcellular Molecular Imaging to Tailor‐Made Therapeutics

Angewandte Chemie International Edition, Journal Year: 2024, Volume and Issue: 63(36)

Published: June 14, 2024

Mitochondria, one of the most important organelles, represent a crucial subcellular target for fundamental research and biomedical applications. Despite significant advances in design DNA nanotechnologies variety bio-applications, dearth strategies that enable mitochondria targeting molecular imaging therapy remains an outstanding challenge this field. In Minireview, we summarize recent progresses on emerging application nanotechnology mitochondria-targeted tumor treatment. We first highlight engineering mitochondria-localized nanosensors situ detection diverse key molecules are essential to maintain mitochondrial functions, including microRNA, enzymes, small molecules, metal ions. Then, compile developments anti-tumor therapy, modularly designed nanodevices delivery therapeutic agents, programmed assembly interference. will place emphasis clarification chemical principles how nanobiotechnology can be various Finally, remaining challenges future directions field discussed, hoping inspire further development advanced toolkits both academic clinical regarding mitochondria.

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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Dual-Locked Enzyme-Activatable Fluorescence Probes for Precise Bioimaging

ACS Biomaterials Science & Engineering, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 22, 2025

Real-time visualization of endogenous enzymes not only helps reveal the underlying biological principles but also provides pathological information for cancer/disease diagnosis and even treatment guidance. To this end, enzyme-activatable fluorescence probes are frequently fabricated that turn their signals "on" exclusively at enzyme-rich region, thus enabling noninvasive real-time imaging interest molecular level with superior sensitivity selectivity. However, in a complex context, commonly used single (i.e., single-locked) may suffer from "false positive" healthy tissues be insufficient to accurately indicate occurrence certain diseases. Therefore, dual-locked have been promoted address these issues. Using dual (or an enzyme another stimulus) as "keys", they permit simultaneous detection distinct biomarkers, offering significantly enhanced precision toward events. Considering recent reviews on remain scarce, we provide review. We summarize progress, particularly highlighting breakthroughs last three years, discuss challenges field.

Language: Английский

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Modular Engineering of Aptamer‐Based Nanobiotechnology for Conditional Control of ATP Sensing

Advanced Materials, Journal Year: 2023, Volume and Issue: 36(22)

Published: Nov. 27, 2023

Dynamic changes of intracellular, extracellular, and subcellular adenosine triphosphates (ATPs) have fundamental interdependence with the physio-pathological states cells. Spatially selective in situ imaging such ATP dynamics offers valuable mechanistic insights into related biological activities. Despite significant advances design aptamer sensors for detection, dearth methods that enable precise specific cellular locations remains a challenge this field. This review focuses on modular engineering regulatable sensing technology via integration probe designs advanced functional nanomaterials, allowing conditional control high spatial precision from organelles to living animals. Highlighting recent photo-triggered nanosensors spatiotemporally controlled imaging, endogenously-triggered cell-selective manner, spatially-controlled nanodevices extracellular microenvironments. Emphasis will be put elucidating principles how nanotechnology can applied regulate aptamer-based The authors envision perspective provides nanobiotechnology opening new frontiers molecular other bio-applications.

Language: Английский

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Modular Weaving DNAzyme in Skeleton of DNA Nanocages for Photoactivatable Catalytic Activity Regulation

Angewandte Chemie, Journal Year: 2024, Volume and Issue: 136(18)

Published: March 22, 2024

Abstract DNAzymes exhibit tremendous application potentials in the field of biosensing and gene regulation due to its unique catalytic function. However, spatiotemporally controlled DNAzyme activity remains a daunting challenge, which may cause nonspecific signal leakage or silencing systems. Here, we report photochemical approach via modular weaving active into skeleton tetrahedral DNA nanocages (TDN) for light‐triggered on‐demand liberation thus conditional control activity. We demonstrate that direct encoding TDN could improve biostability ensure delivery efficiency, comparing with conventional surface anchoring strategy. Furthermore, molecular nanostructures allows remote DNAzyme‐mediated high spatiotemporal precision light. In addition, is applicable editing functions other functional nucleic acids.

Language: Английский

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