Chemical Engineering of DNAzyme for Effective Biosensing and Gene Therapy

Small Methods, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 2, 2025

RNA-cleaving DNAzymes are in vitro selected functional nucleic acids with inherent catalytic activities. Due to their unique properties, such as high specificity, substrate cleavage capability, and programmability, have emerged powerful tools the fields of analytical chemistry, chemical biology, biomedicine. Nevertheless, biological applications still impeded by several challenges, structural instability, compromised activity environments lack spatiotemporal control designs, which may result false-positive signals, limited efficacy or non-specific activation associated side effects. To address these various strategies been explored regulate DNAzyme through modifications, enhancing stability, selectivity, functionality, thereby positioning them ideal candidates for applications. In this review, a comprehensive overview chemically modified is provided, discussing modification effects modifications on DNAzymes. Specific examples use biosensing gene therapy also presented discussed. Finally, current challenges field addressed offer perspectives potential direction

Language: Английский

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Endogenous Glutathione-Activated Nucleic Acid Molecular Circuitry for Cell-Specific MicroRNA Imaging

Analytical Chemistry, Journal Year: 2024, Volume and Issue: 96(31), P. 12854 - 12861

Published: July 23, 2024

Sensitive and reliable microRNA imaging in living cells has significant implications for clinical diagnosis monitoring. Catalytic DNA circuits have emerged as potent tools tracking these intracellular biomarkers probing the corresponding biochemical processes. However, their utility is hindered by low resistance to external interference, leading undesired off-site activation consequent signal leakage. Therefore, achieving endogenous control of circuit's preferable target analysis cells. In this study, we attempted address challenge engineering a simple yet effective glutathione (GSH)-regulated hybridization chain reaction (HCR) circuit acquiring high-contrast miRNA imaging. Initially, HCR hairpin reactants were blocked engineered disulfide-integrated duplex, thus effectively passivating sensing function. And precaged was liberated cell-specific GSH molecule, initiating system selectively amplified detection microRNA-21 (miR-21). This approach prevented unwanted leakage before exposure into cells, ensuring robust miR-21 with high accuracy reliability specific tumor Moreover, endogenously responsive established link between small regulatory factors miRNA, thereby enhancing gain. summary, activatable represents versatile toolbox bioanalysis exploration potential signaling pathways

Language: Английский

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A Trinity DNAzyme Circuit with Dual‐Confinement Effect for Imaging‐Guided Combined Gene/Chemo‐Dynamic Therapy

Advanced Functional Materials, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 23, 2025

Abstract Synthetic catalytic DNAzyme circuits have gained great attention as versatile theranostic toolboxes, but their accuracy and efficiency is restricted by uncontrolled signal leakage inefficient circuitry activation. Here, a trinity circuit (TriDC) constructed using unique logic gate bioswitchable DNA tetrahedral walker@MnO 2 nanosheets structure. The AND walker, composed of dual allosteric (termed “commander” “soldier”) scaffold, used to execute imaging‐guided gene therapy avoiding undesired cascade “walking” in confined space. Glutathione‐mediated reduction MnO provides secondary confinement stack the walker situ, offering abundant Mn 2+ cofactor achieve self‐sufficiency induce chemo‐dynamic therapy. structure increases local reaction concentrations ≈100‐fold according collision frequency model, enabling detection microRNA‐10b microRNA‐155 at limit 98.97 p m , thereby facilitating precise imaging breast cancer cells. combined gene/chemo‐dynamic achieves 66.2% tumor growth inhibition efficiency. TriDC offers novel insights into theranostics also presents paradigm for situ implementation higher‐order circuits.

Language: Английский

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Leveraging Demethylase Activation in DNA Circuits to Overcome Signal Leakage for Reliable MicroRNA Bioimaging

Small, Journal Year: 2025, Volume and Issue: unknown

Published: April 10, 2025

Abstract DNA circuits show great potential in monitoring intracellular biomarkers based on their high programmability, predictability, and unique signal amplification capabilities, yet face challenges from uncontrollable leakage caused by the complex environment. Herein, a demethylase‐activated DNA‐assembly (DAD) circuit is designed for reliable robust imaging of cellular microRNA, incorporating sequential activation hybridization chain reaction (HCR) amplifier system. The DAD consists DNAzyme module microRNA‐recognizing HCR signal‐amplifying module. m 6 A‐modified sequence module, initially possessing temporally caged substrate‐cleavage activity, integrated into probe effectively blocking its miRNA‐sensing capacity. In presence ALKBH5 demethylase, methyl‐modifying unit removed, thus restoring catalytic substrate‐cleaving activity. This process exposed previously toehold region probe, thereby activating sensing miRNA. By leveraging activation, this can substantially enhance signal‐to‐background ratio, enabling highly sensitive miRNA detection efficient differentiation cancerous normal cells. Furthermore, established relationship between demethylase enzyme miRNA, paving way investigating more complicate biological processes intricate signaling pathways within

Language: Английский

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Sequentially amplified integration of catalytic DNA circuits for high-performance intracellular imaging of miRNA and interpretation of mRNA-miRNA signalling pathway

Nano Today, Journal Year: 2024, Volume and Issue: 61, P. 102563 - 102563

Published: Dec. 1, 2024

Language: Английский

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Redox-Stimulated Catalytic DNA Circuit for High-fidelity Imaging of MicroRNA and in situ Interpretation of the Relevant Regulatory Pathway

Biosensors and Bioelectronics, Journal Year: 2024, Volume and Issue: 272, P. 117109 - 117109

Published: Dec. 28, 2024

Language: Английский

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A Methyl‐Engineered DNAzyme for Endogenous Alkyltransferase Monitoring and Self‐Sufficient Gene Regulation

Small Methods, Journal Year: 2024, Volume and Issue: unknown

Published: Sept. 19, 2024

Abstract The on‐demand gene regulation is crucial for extensively exploring specific functions and developing personalized therapeutics, which shows great promise in precision medicines. Although some nucleic acid‐based regulatory tools (antisense oligonucleotides small interfering RNAs) are devised achieving activation, the introduction of chemical modifications may cause undesired side effects, thereby impairing efficacy. Herein, a methyl‐engineered DNAzyme (MeDz) developed visualization endogenous alkyltransferase (AGT) simultaneous self‐sufficiently regulation. catalytic activity can be efficiently blocked by O 6 ‐methylguanine (O MeG) modification specifically restored via AGT‐mediated DNA‐repairing pathway. This simply designed MeDz demonstrated to reveal AGT varying expression levels different cells, opening possibility explore AGT‐related biological processes. Moreover, AGT‐guided exhibits cell‐selective on human early growth response‐1 (EGR‐1) gene, with efficient repression breast cancer cells low effectiveness normal cells. proposed offers an attractive strategy regulation, displaying potential biomedical applications.

Language: Английский

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Self‐Adaptive Activation of DNAzyme Nanoassembly for Synergistically Combined Gene Therapy

Angewandte Chemie, Journal Year: 2024, Volume and Issue: unknown

Published: Oct. 17, 2024

Abstract DNAzyme represents a promising gene silencing toolbox yet is obstructed by the poor substrate accessibility in specific cells. Herein, compact DNA nanoassembly, incorporating multimeric therapeutic DNAzyme, was prepared for selective delivery of gene‐silencing with requisite cofactors and auxiliary chemo‐drugs. By virtue sequence‐conservative duplex‐specific nuclease, endogenous miRNA catalyzes successive site‐specific cleavage nanoassembly (nominated as localized RNA walking machine) thus ensures liberation/activation agents high accuracy efficacy. The miR‐10b‐stimulated designed to downregulate TWIST transcription factor, an upstream promotor miR‐10b, acquiring self‐sufficient downregulation TWIST/miR‐10b signaling nodes (self‐adaptive negative feedback loop) abrogating tumor metastasis chemo‐resistance issues.

Language: Английский

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Harnessing Demethylase-Regulated Catalytic DNA Circuit for In-Situ Investigation of the Regulatory Connection with MicroRNA

Analytical Chemistry, Journal Year: 2024, Volume and Issue: 96(51), P. 20304 - 20311

Published: Dec. 12, 2024

Insight into the epigenetic modulation-correlated molecule interactions has significant implications for in-depth understanding of intracellular intricate biological networks. However, there is currently a lack reliable tools elucidating potential correlation between regulators and relevant genes, e.g., microRNAs (miRNAs). Herein, an alkB homologue 5 (ALKBH5, key regulator)-modulated catalytic DNA circuit (ACD) was constructed by grafting N6-methyladenosine (m6A)-caged I-R3 DNAzyme circuitry components achieving on-site miRNA imaging in living cells. Specifically, activity could be effectively suppressed m6A modification situated at its highly sequence-conserved core region then selectively restored through ALKBH5-mediated demethylation pathway. And ALKBH5-activated allowed efficient cleaving reaction presence cofactors, resulting liberation hairpin assembly (CHA) reactants. Subsequently, target triggered CHA to produce duplex product while releasing analyte. The liberated autonomously trigger next round cycle generating amplified fluorescence readout. By virtue stimuli-responsive activation amplification circuit, ACD system achieved specific sensitive tumor Moreover, this efficiently reliably demonstrated reveal underlying relationship activator ALKBH5 miRNA. Overall, developed provides promising tool robust profiling epigenetic-involved signal pathways, thus displaying great bioanalytical applications.

Language: Английский

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Engineering of a DNAzyme-based aptasensor for highly efficient ATP detection and oyster freshness evaluation

Journal of Food Composition and Analysis, Journal Year: 2024, Volume and Issue: unknown, P. 107176 - 107176

Published: Dec. 1, 2024

Language: Английский

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