Engineering DNA Nanodevices with Multi-site Recognition and Multi-signal Output for Accurate Intracellular LncRNA Imaging

Analytical Chemistry, Год журнала: 2025, Номер unknown

Опубликована: Фев. 5, 2025

Dynamic DNA nanodevices, known for their high programmability and controllability, are pivotal in intracellular biomarker imaging. However, these nanodevices often suffer from inadequate detection sensitivity specificity due to limited cellular loading capacity low signal feedback. Herein, we engineered an integrated multi-site recognition multi-signal output of four-leaf clover dynamic nanodevice (MEMORY) that enables sensitive accurate long noncoding RNA (lncRNA) MEMORY features one fluorophore (FAM)-modified cross-shaped structure as spatial-confinement scaffolds loaded with four identical quenchers (BHQ1)-modified probes (RPs), ensuring a background initially. In the presence target lncRNA, multiple sites facilitate hybridization selectively release RPs, exposing toehold region outputting green fluorescence (FAM) signal. Furthermore, exposed can trigger efficient rapid chain reaction (HCR) amplification, red (Cy5) MEMORY's increase likelihood collisions, enhancing efficiency, while its provides sequential feedback through FAM Cy5, boosting overall intensity. With lncRNA metastasis-related lung adenocarcinoma transcript 1 (MALAT1) model, offers linear range pM 100 nM, limit 0.29 pM. We demonstrated differentiate between normal tumor cells based on MALAT1 This will offer valuable tools imaging biomarkers.

Язык: Английский

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Programmable Split DNAzyme Modulators via Allosteric Cooperative Activation for mRNA Electrochemiluminescence Biosensing

Analytical Chemistry, Год журнала: 2025, Номер unknown

Опубликована: Март 3, 2025

DNAzymes, known for their programmability, stability, and cost-effectiveness, are powerful tools signal transduction in complex biological systems. However, application responding to target effectors is often hindered by limited catalytic efficiency susceptibility unintended activation. Here we propose an allosteric cooperative activation strategy program a split DNAzyme modulator (STATER) that enables sensitive accurate electrochemiluminescence (ECL) biosensing of interleukin-6 (IL-6) mRNA. Our design features STATER leverages DNA tetrahedron as central scaffold, equipped with two pairs T-shaped hairpin probes (TP) helper (HP). Specifically, the TP contains apurinic/apyrimidinic endonuclease 1 (APE1) recognition sites, IL-6 mRNA region, partzyme fragment, while HP corresponding paired fragment. Unlike conventional modulators rely on single effector activation, integrates mechanism, which ensures all preblocked components synergistically activated assembled within confined space, facilitating rapid specific reconstruction DNAzyme's active domain. Furthermore, upon APE1 mRNA, inactive partzymes undergo assembly via toehold exchange displacement reaction, switching cleavage reactivity STATER. This mechanism establishment threshold thereby minimizing nonspecific scenarios. studies demonstrate exhibits outstanding sensitivity selectivity detection using supramolecular gold nanoclusters network-based ECL platform. The biosensor provides linear span from × 10–13 10–7 M, limit low 3.26 10–14 highlighting STATER's potential detecting various analytes

Язык: Английский

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Endogenous Enzyme-Activated Spatial Confinement DNA Nanowire with a Tumor Cell-Specific Response for High-Precision Imaging of the Tumor/Normal Cells Boundary

Analytical Chemistry, Год журнала: 2025, Номер unknown

Опубликована: Апрель 12, 2025

Developing tumor cell-specific imaging approaches is essential for the clear delineation of margins. However, traditional suffered from low reaction kinetics as well limited specificity resulting their "always active" sensing mode, making it difficult to accurately depict boundary. To address these limitations, we developed an endogenous enzyme-activated spatial confinement DNA nanowire probe (E-SCNW) with enhanced tumor/normal cell discrimination ratio high precision cells The effect can improve kinetics, and enzyme-activation design confine fluorescence response region. Additionally, no additional delivery carriers were required during cross membrane into intracellular space. It worth noting that benefiting design, detection limit was decreased by nearly 25.6-fold 4.46-fold through using E-SCNW, indicating promising prospects in high-precision

Язык: Английский

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Specific Response Assembly of 3D Space-Confined DNA Nanoaggregates for Rapid and Sensitive Detection of DNA Methyltransferase

Analytical Chemistry, Год журнала: 2024, Номер unknown

Опубликована: Дек. 18, 2024

Rapid and sensitive detection of DNA adenine methyltransferase (Dam) activity is crucial for both research clinical applications. Herein, we utilize two types spherical nucleic acids (SNAs) to specific response assemble into 3D space-confined nanoaggregates that enable the rapid Dam activity. The SNAs feature order scaffolds serve as cores anchoring signal hairpin probes (S-HPs) target (T-HPs). Specifically, distinct S-HPs are labeled with FAM fluorophores BHQ1 quenchers share identical sequences, while T-HPs designed different linking sequences recognition regions, resulting in formation (SNA1 SNA2). In presence Dam, region methylated subsequently cleaved by auxiliary endonuclease, releasing loop T-HP a walking strand exposing sequence on SNAs. Notably, prior design complementary facilitates their assembly nanoaggregates, creating confined space strands recover fluorescent signals. nanoaggregate system not only provides highly ordered tracks but also enhances spatial continuity strands, greatly improving reaction kinetics detecting This strategy enables within 105 min, achieving limit 2.9 × 10

Язык: Английский

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