Duplex-Specific Nuclease-Assisted CRISPR-Cas12a Strategy for MicroRNA Detection Using a Personal Glucose Meter DOI
Shaohua Gong, Jingjing Li, Wei Pan

и другие.

Analytical Chemistry, Год журнала: 2021, Номер 93(30), С. 10719 - 10726

Опубликована: Июль 19, 2021

A CRISPR-Cas system holds great promise as a next-generation biosensing technology for molecular diagnostics. Nevertheless, the current CRISPR-Cas12a-based detection strategies always need bulky instruments or auxiliary devices to obtain quantitative signal output, which restrains its point-of-care testing application. Herein, we proposed duplex-specific nuclease-assisted CRISPR-Cas12a strategy detect microRNA (miRNA) with personal glucose meter. The target miRNA was first converted into an amplified initiator DNA via nuclease. Afterward, activated collateral cleavage activity of cleave single-strand (ssDNA) linker on sucrase-ssDNA-modified magnetic beads, led release sucrase. released sucrase collected and then utilized catalyze sucrose glucose, could be quantitatively detected by change in directly reflected concentration miRNA, avoided expensive equipment quantification. Two different miRNAs (miRNA21 miRNA205) simply changing sequence template strand (H strand). developed showed high sensitivity limit (LOD) 2.4 1.1 pM miRNA21 miRNA205, respectively. In addition, good selectivity anti-interference ability were achieved using this method, enabled it promising at point-of-care.

Язык: Английский

Controllable Assembly of a Quantum Dot-Based Aptasensor Guided by CRISPR/Cas12a for Direct Measurement of Circulating Tumor Cells in Human Blood DOI
Qian Zhang, Xin Gao, Yi‐Ping Ho

и другие.

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

Процитировано

33
CRISPR-Based Homogeneous Electrochemical Strategy for Near-Zero Background Detection of Breast Cancer Extracellular Vesicles via Fluidity-Enhanced Magnetic Capture Nanoprobe DOI
Limin Yang, Jingang Zhang, Jing Zhang

и другие.

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

Процитировано

6
Micropore Resistance Counting Platform for Multiplexed and Ultrasensitive Detection of Mycotoxins and Biomarkers DOI
Letian Li, Mengjiao Wang, Yongzhen Dong

и другие.

ACS Nano, Год журнала: 2025, Номер unknown

Опубликована: Янв. 3, 2025

Development of a multiplexed and sensitive biosensing platform is priority for public health security. We report micropore resistance counting based on polystyrene microsphere size-based encoding Clostridium butyricum Argonaute (CbAgo) decoding ultrasensitive detection. Initially, we constructed target DNA-modified coding system counting. Subsequently, the precise recognition cleavage capabilities guide DNA-activated CbAgo protein enable encoded system. Changes in concentration microspheres are presented as signal readout. The demonstrated excellent performance detection three mycotoxins (with sensitivity range over 4 orders magnitude reaching pg/mL level) two inflammatory markers at pg/mL. Combining enzyme by with counting, developed highly tool wide-ranging potential applications such clinical diagnosis, food safety inspection, environmental monitoring.

Язык: Английский

Процитировано

5
Dual miRNAs Imaging Platform Based on HRCA-Cas12a by Replacing PAM with Bubble to Reduce False Positive DOI
Xiaojuan Wang, Chenxi Zhao,

Nana Yin

и другие.

Analytical Chemistry, Год журнала: 2025, Номер unknown

Опубликована: Янв. 29, 2025

Detection and imaging of dual miRNAs based on AND logic gates can improve the accuracy early diagnosis disease. However, a single target may lead to false positive. Hence, this work rationally integrates hyperbranched rolling circle amplification (HRCA) with Cas12a by replacing PAM sequence bubble sensitively detect image miRNA-10b miRNA-21 gate. When are both present, two padlocks linked into circular DNA as template for RCA. Long ssDNA products generated under catalysis phi29 polymerase, which cis-cleaved activated trans-cleavage generate fluorescent signals. Subsequently, primer hybridizes cis-cleavage is extended dsDNA substrate produce more produces significant signals leading positive due presence protospacer adjacent motif (PAM) padlock. After removed from padlock, RCA form bubbles replace PAM, activate without affecting sensitivity reduce The introduction enables second utilization Cas12a, increasing signal-to-noise ratio. HRCA exhibit optimal activity in T4 ligase buffer, achieving one-pot detection miRNAs. In addition, HRCA-Cas12a method intracellular visualization It exhibits ability distinguish different types cancer cells expression level

Язык: Английский

Процитировано

3
Duplex-Specific Nuclease-Assisted CRISPR-Cas12a Strategy for MicroRNA Detection Using a Personal Glucose Meter DOI
Shaohua Gong, Jingjing Li, Wei Pan

и другие.

Analytical Chemistry, Год журнала: 2021, Номер 93(30), С. 10719 - 10726

Опубликована: Июль 19, 2021

A CRISPR-Cas system holds great promise as a next-generation biosensing technology for molecular diagnostics. Nevertheless, the current CRISPR-Cas12a-based detection strategies always need bulky instruments or auxiliary devices to obtain quantitative signal output, which restrains its point-of-care testing application. Herein, we proposed duplex-specific nuclease-assisted CRISPR-Cas12a strategy detect microRNA (miRNA) with personal glucose meter. The target miRNA was first converted into an amplified initiator DNA via nuclease. Afterward, activated collateral cleavage activity of cleave single-strand (ssDNA) linker on sucrase-ssDNA-modified magnetic beads, led release sucrase. released sucrase collected and then utilized catalyze sucrose glucose, could be quantitatively detected by change in directly reflected concentration miRNA, avoided expensive equipment quantification. Two different miRNAs (miRNA21 miRNA205) simply changing sequence template strand (H strand). developed showed high sensitivity limit (LOD) 2.4 1.1 pM miRNA21 miRNA205, respectively. In addition, good selectivity anti-interference ability were achieved using this method, enabled it promising at point-of-care.

Язык: Английский

Процитировано

99