Programmable CRISPR-mediated gold nanoparticle adhesion for visual colorimetric detection

Dec 17, 2025Biosensors & bioelectronics

Using CRISPR-controlled gold nanoparticles to detect substances by color change

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Abstract

Detection of human papillomavirus (HPV) DNA was achieved down to 10 aM using a novel colorimetric assay.

A programmable adhesion phenomenon was created using gold nanoparticles functionalized with specific DNA probes.
Localized aggregation and strong adhesion to hydrophobic surfaces were induced by hydrophobic components on the nanoparticle surface.
The assay utilizes CRISPR-Cas12a-mediated DNA cleavage for visual detection of nucleic acids.
Combining this method with recombinase polymerase amplification enhances sensitivity for HPV DNA detection.
The platform was validated with cervical swab samples, indicating potential for low-cost diagnostics.

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While the aggregation behavior of gold nanoparticles (AuNPs) has been extensively studied in biosensing, catalysis, and nanomedicine, their potential for programmable surface adhesion via tunable surface chemistry remains largely untapped. Here, a programmable CRISPR-mediated hydrophobic adhesion phenomenon using streptavidin-coated AuNPs functionalized with Cy5-ssDNA-biotin probes is introduced. Hydrophobic Cy5 moieties on the AuNP surface induce localized aggregation and strong adhesion to hydrophobic surfaces. This unique behavior was leveraged by coupling CRISPR-Cas12a-mediated ssDNA cleavage with Cy5-labeled ssDNA-coated AuNPs to develop a simple, visual-readout colorimetric assay for nucleic acid detection. When combined with recombinase polymerase amplification, the method achieved ultrasensitive detection of human papillomavirus (HPV) DNA down to 10 aM, without the need for complex instrumentation. The platform's clinical utility was validated by detecting HPV DNA in cervical swab samples, highlighting its promise for low-cost, sensitive, and accessible point-of-care diagnostics in resource-limited settings. Unlike conventional aggregation-dispersion systems, this platform introduces a fundamentally distinct signal transduction mechanism based on surface adhesion, defining a new modality within CRISPR-based colorimetric diagnostics and offering a simple, low-cost solution for point-of-care testing.

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Programmable CRISPR-mediated gold nanoparticle adhesion for visual colorimetric detection

Abstract

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