Filter Membrane-Based Colorimetric Approach for Point-of-Care Detection of Biomarkers Using CRISPR-Cas12a

Analytical Chemistry, Journal Year: 2024, Volume and Issue: 96(39), P. 15789 - 15796

Published: Sept. 23, 2024

CRISPR-Cas-based point-of-care testing (POCT) strategies have been widely explored for the detection of diverse biomarkers. However, these methods often require complicated operations, such as careful solution transfer steps, to achieve high sensitivity and accuracy. In this study, we combine a filter membrane-based POCT method with CRISPR-Cas12a colorimetric For nucleic acid target, trans-cleavage activity is directly triggered, cutting single-stranded DNA linkers on glucose oxidase (GOx)-modified polymer nanoparticles. Due size difference between GOx nanoparticles, can be separated using membrane. The filtrate containing reacts substrate generate signal. non-nucleic signal converted into that activates CRISPR-Cas12a, resulting in entire operation easy perform, observed via naked eye, which circumvents use costly instruments. developed strategy holds great promise accurate accessible disease biomarkers resource-limited settings.

Language: Английский

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Phosphorothioate-Modified Hairpin G-Triplex Reporter-Assisted Split CRISPR/Cas12a-Powered Biosensor for “Turn-On” Fluorescent Detection of Nucleic Acid and Non-Nucleic Acid Targets

Analytical Chemistry, Journal Year: 2025, Volume and Issue: unknown

Published: April 24, 2025

CRISPR/Cas12a-powered biosensors with guanine (G)-rich sequence reporters (e.g., G-quadruplex and G-triplex) are widely used in detection applications due to their simplicity sensitivity. However, when these employed for molecular complex samples, they may encounter difficulties such as high background signal susceptibility interference because of the "turn-off" output. Herein, we explore, first time, a set phosphorothioate (ps)-modified (G4) G-triplex (G3) sequences that can bind thioflavin T (ThT) an active split CRISPR/Cas12a system (SCas12a) generate "turn-on" fluorescent signal. To apply this new phenomenon, develop universal SCas12a-powered biosensor nucleic acid (miRNA-21) non-nucleic (kanamycin) targets by using ps-modified hairpin G3 reporter (SCas12a/psHG3). Target recognition activates SCas12a's trans-cleavage activity, leading cleavage at loop region psHG3 reporter. The released prelocked psG3 DNA binds ThT produce strong fluorescence Without preamplification, strategy detect miRNA-21 limit 100 fM. Moreover, SCas12a/psHG3 was further utilized detecting kanamycin incorporating its aptamers, enabling concentrations low pM. This work is system, showcasing improved performance wide range synthetic biology-based sensing technology.

Language: Английский

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Aptamer-based dual-enzyme, amplification-free biosensor integrating CRISPR-Cas12a and Exo III for sensitive detection of ATP

Sensing and Bio-Sensing Research, Journal Year: 2025, Volume and Issue: unknown, P. 100795 - 100795

Published: April 1, 2025

Language: Английский

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CRISPR/Cas12a dual-mode biosensor for Staphylococcus aureus detection via enzyme-free isothermal amplification

Talanta, Journal Year: 2024, Volume and Issue: 282, P. 127013 - 127013

Published: Oct. 10, 2024

Language: Английский

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One-pot method based on CRISPR/Cas12a-triggered entropy-driven RNA circuit for sensitive glycated hemoglobin A1c assay

Sensors and Actuators B Chemical, Journal Year: 2024, Volume and Issue: unknown, P. 136720 - 136720

Published: Oct. 1, 2024

Language: Английский

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Asymmetric CRISPR-Cas12a powered electrochemical aptasensor for clenbuterol detection based on competitive gRNA mediated cascade signal amplification

Food Chemistry, Journal Year: 2024, Volume and Issue: 464, P. 141928 - 141928

Published: Nov. 4, 2024

Language: Английский

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