Controllable Assembly of a Quantum Dot-Based Aptasensor Guided by CRISPR/Cas12a for Direct Measurement of Circulating Tumor Cells in Human Blood

Nano Letters, Год журнала: 2024, Номер 24(7), С. 2360 - 2368

Опубликована: Фев. 12, 2024

Accurate and sensitive analysis of circulating tumor cells (CTCs) in human blood provides a non-invasive approach for the evaluation cancer metastasis early diagnosis. Herein, we demonstrate controllable assembly quantum dot (QD)-based aptasensor guided by CRISPR/Cas12a direct measurement CTCs blood. We introduce magnetic bead@activator/recognizer duplex core–shell structure to construct multifunctional platform capture detection blood, without need additional CTC release re-identification steps. Notably, introduction separation ensures that only target-induced free activator can initiate downstream catalysis, efficiently avoiding undesired catalysis triggered inappropriate recognition activator/recognizer crRNAs. This achieves high CTC-capture efficiency (82.72%) with limit 2 mL–1 holding great promise liquid biopsy cancers.

Язык: Английский

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Dual-microRNA-Controlled Electrochemiluminescence Biosensor for Breast Cancer Diagnosis and Supplemental Identification of Breast Cancer Metastasis

Analytical Chemistry, Год журнала: 2024, Номер 96(8), С. 3636 - 3644

Опубликована: Фев. 15, 2024

Breast cancer remains the most frequently diagnosed globally, and metastasis of this malignancy is primary cause mortality in breast patients. Hence, prompt diagnosis timely detection metastatic are critical for effective therapeutic intervention. Both progression closely associated with aberrant expression specific microRNAs (miRNAs) enzymes. To facilitate concomitant identification cancer, we have engineered an innovative electrochemiluminescence (ECL)-based sensing platform integrated enzyme-free DNA amplification circuits dual functionality. Specifically, microRNA-21 (miR-21) employed as a biomarker miR-21 induces quenching ECL signal from luminophores via strategically designed catalytic three-hairpin assembly (CTHA) circuit. Subsequently, miR-105 levels measured toehold-mediated strand displacement reactions (TSDR). Here, restores initially quenched signal, enabling assessment propensity. Our experimental data demonstrate that devised biosensor offers broad linear ranges low limits both miR-105. Importantly, our novel was also successfully validated by using cellular serum samples. This not only discriminates cell lines MCF-7 MDA-MB-231 nonbreast cells like HepG2, TPC-1, HeLa, but it distinguishes between malignant cells. Consequently, approach holds significant promise clinical applications precise screening.

Язык: Английский

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

Biosensors and Bioelectronics, Год журнала: 2024, Номер 250, С. 116060 - 116060

Опубликована: Янв. 23, 2024

Язык: Английский

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Buoyant Metal–Organic Framework Corona-Driven Fast Isolation and Ultrasensitive Profiling of Circulating Extracellular Vesicles

ACS Nano, Год журнала: 2024, Номер 18(22), С. 14569 - 14582

Опубликована: Май 23, 2024

Accurately assaying tumor-derived circulating extracellular vesicles (EVs) is fundamental in noninvasive cancer diagnosis and therapeutic monitoring but limited by challenges efficient EV isolation profiling. Here, we report a bioinspired buoyancy-driven metal-organic framework (MOF) corona that leverages on-bubble coordination dual-encoded surface-enhanced Raman scattering (SERS) nanotags to streamline rapid ultrasensitive profiling of plasma EVs single assay for diagnostics. This integrated bubble-MOF-SERS (IBMsv) allows barnacle-like high-density adhesion MOFs on self-floating bubble surface enable fast (2 min, near 90% capture efficiency) tumor via enhanced EV-MOF binding. Also, IBMsv harnesses four-plexed SERS profile the captured protein markers at single-particle level. Such sensitive multiplexed across five types, revealing heterogeneous expression patterns. Furthermore, enables pilot clinical cohort (

Язык: Английский

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CRISPR-Based Homogeneous Electrochemical Strategy for Near-Zero Background Detection of Breast Cancer Extracellular Vesicles via Fluidity-Enhanced Magnetic Capture Nanoprobe

Analytical Chemistry, Год журнала: 2025, Номер unknown

Опубликована: Янв. 19, 2025

Precise identification and analysis of multiple protein biomarkers on the surface breast cancer cell-derived extracellular vesicles (BC-EVs) are great significance for noninvasive diagnosis subtypes, but it remains a major challenge owing to their high heterogeneity low abundance. Herein, we established CRISPR-based homogeneous electrochemical strategy near-zero background ultrasensitive detection BC-EVs. To realize high-performance capture isolation BC-EVs, fluidity-enhanced magnetic nanoprobes were facilely prepared. After capturing AND logic gate-based catalytic hairpin assembly (CHA) trans-cleavage activity CRISPR-Cas12a against signal triggered successively, generating significant signal. Notably, as-developed metal-mediated could efficiently decrease by separation, endowing method with signal-to-noise ratio. Consequently, ingeniously integrating DNA CRISPR-CHA amplification dual in strategy, precise BC-EVs was successfully achieved through simultaneous specific recognition markers surface. More importantly, this approach effectively discriminate subgroups clinical serum samples, which may provide opportunities accurate prognosis evaluation manner.

Язык: Английский

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Simultaneous subset tracing and miRNA profiling of tumor-derived exosomes via dual-surface-protein orthogonal barcoding

Science Advances, Год журнала: 2023, Номер 9(40)

Опубликована: Окт. 4, 2023

The clinical potential of miRNA-based liquid biopsy has been largely limited by the heterogeneous sources in plasma and tedious assay processes. Here, we develop a precise robust one-pot called dual-surface-protein-guided orthogonal recognition tumor-derived exosomes situ profiling microRNAs (SORTER) to detect exosomal miRNAs enhance diagnostic accuracy prostate cancer (PCa). SORTER uses two allosteric aptamers against marker CD63 tumor EpCAM create an labeling barcode achieve selective sorting tumor-specific exosome subtypes. Furthermore, labeled on initiated targeted membrane fusion with liposome probes import miRNA detection reagents, enabling sensitive miRNAs. With signature six miRNAs, differentiated PCa benign prostatic hyperplasia 100%. Notably, reached 90.6% classification metastatic nonmetastatic PCa. We envision that will promote adaptability biopsy.

Язык: Английский

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An Integrated Amplification-Free Digital CRISPR/Cas-Assisted Assay for Single Molecule Detection of RNA

ACS Nano, Год журнала: 2023, Номер 17(8), С. 7250 - 7256

Опубликована: Апрель 13, 2023

Conventional nucleic acid detection technologies usually rely on amplification to improve sensitivity, which has drawbacks, such as bias, complicated operation, high requirements for complex instruments, and aerosol pollution. To address these concerns, we developed an integrated assay the enrichment single molecule digital of based a CRISPR/Cas13a microwell array. In our design, magnetic beads capture concentrate target from large volume sample, is 100 times larger than reported earlier. The target-induced cutting reaction was then dispersed limited million individual femtoliter-sized microwells, thereby enhancing local signal intensity achieve single-molecule detection. limit this amplification-free SARS-CoV-2 2 aM. implementation study will establish "sample-in-answer-out" single-RNA technology without sensitivity specificity while shortening time. This research broad prospects in clinical application.

Язык: Английский

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Selective In Situ Analysis of Mature microRNAs in Extracellular Vesicles Using a DNA Cage‐Based Thermophoretic Assay

Angewandte Chemie International Edition, Год журнала: 2023, Номер 62(24)

Опубликована: Апрель 20, 2023

Abstract Mature microRNAs (miRNAs) in extracellular vesicles (EVs) are involved different stages of cancer progression, yet it remains challenging to precisely detect mature miRNAs EVs due the presence interfering RNAs (such as longer precursor miRNAs, pre‐miRNAs) and low abundance tumor‐associated miRNAs. By leveraging size‐selective ability DNA cages polyethylene glycol (PEG)‐enhanced thermophoretic accumulation EVs, we devised a cage‐based assay for highly sensitive, selective, situ detection with limit (LoD) 2.05 fM. Our can profile EV directly serum samples without interference pre‐miRNAs need ultracentrifugation. A clinical study showed that miR‐21 or miR‐155 had an overall accuracy 90 % discrimination between breast patients healthy donors, which outperformed conventional molecular probes detecting both pre‐miRNAs. We envision our advance miRNA‐based diagnosis cancer.

Язык: Английский

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Beyond Extracellular Vesicles: Hybrid Membrane Nanovesicles as Emerging Advanced Tools for Biomedical Applications

Advanced Science, Год журнала: 2023, Номер 10(32)

Опубликована: Сен. 25, 2023

Extracellular vesicles (EVs), involved in essential physiological and pathological processes of the organism, have emerged as powerful tools for disease treatment owing to their unique natural biological characteristics artificially acquired advantages. However, limited targeting ability, insufficient production yield, low drug-loading capability simplex EVs greatly hindered development clinical translation. Therefore, establishment multifunctional hybrid membrane nanovesicles (HMNVs) with favorable adaptability flexibility has become key expanding practical application EVs. This timely review summarizes current progress HMNVs biomedical applications. Different preparation strategies including physical, chemical, chimera approaches are first discussed. then individually describes diverse types based on homologous or heterologous cell substances, a fusion liposome, well bacterial membrane. Subsequently, specific emphasis is placed highlight applications toward various diseases representative examples. Finally, ongoing challenges prospects currently developed translational briefly presented. will not only stimulate broad interest among researchers from disciplines but also provide valuable insights promising nanoplatforms precision medicine.

Язык: Английский

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Asymmetrically split DNAzyme-based colorimetric and electrochemical dual-modal biosensor for detection of breast cancer exosomal surface proteins

Biosensors and Bioelectronics, Год журнала: 2023, Номер 238, С. 115552 - 115552

Опубликована: Июль 26, 2023

Язык: Английский

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