CRISPR/Cas9-Based Lateral Flow and Fluorescence Diagnostics

Osborn, Mark J. and Bhardwaj, Akshay and Bingea, Samuel P. and Knipping, Friederike and Feser, Colby J. and Lees, Christopher J. and Collins, Daniel P. and Steer, Clifford J. and Blazar, Bruce R. and Tolar, Jakub (2021) CRISPR/Cas9-Based Lateral Flow and Fluorescence Diagnostics. Bioengineering, 8 (2). p. 23. ISSN 2306-5354

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Abstract

Clustered regularly interspaced short palindromic repeat (CRISPR/Cas) proteins can be designed to bind specified DNA and RNA sequences and hold great promise for the accurate detection of nucleic acids for diagnostics. We integrated commercially available reagents into a CRISPR/Cas9-based lateral flow assay that can detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) sequences with single-base specificity. This approach requires minimal equipment and represents a simplified platform for field-based deployment. We also developed a rapid, multiplex fluorescence CRISPR/Cas9 nuclease cleavage assay capable of detecting and differentiating SARS-CoV-2, influenza A and B, and respiratory syncytial virus in a single reaction. Our findings provide proof-of-principle for CRISPR/Cas9 point-of-care diagnosis as well as a scalable fluorescent platform for identifying respiratory viral pathogens with overlapping symptomology.

Item Type: Article
Subjects: Science Repository > Engineering
Depositing User: Managing Editor
Date Deposited: 08 Mar 2023 07:17
Last Modified: 25 May 2024 07:30
URI: http://research.manuscritpub.com/id/eprint/633

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