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Article

CRISPR/Cas9-Based Lateral Flow and Fluorescence Diagnostics

1
Department of Pediatrics, Division of Blood and Marrow Transplant & Cellular Therapy, University of Minnesota Medical School, Minneapolis, MN 55455, USA
2
Cytomedical Design Group, LLC, Saint Paul, MN 55127, USA
3
Department of Medicine, University of Minnesota Medical School, Minneapolis, MN 55455, USA
4
Department of Genetics, Cell Biology and Development, University of Minnesota Medical School, Minneapolis, MN 55455, USA
*
Author to whom correspondence should be addressed.
Academic Editor: Haridha Shivram
Bioengineering 2021, 8(2), 23; https://doi.org/10.3390/bioengineering8020023
Received: 30 December 2020 / Revised: 6 February 2021 / Accepted: 9 February 2021 / Published: 12 February 2021
(This article belongs to the Special Issue CRISPR-Cas: Discovery, Function and Application)
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. View Full-Text
Keywords: CRISPR/Cas9; SARS-Co-V2; lateral flow assay CRISPR/Cas9; SARS-Co-V2; lateral flow assay
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MDPI and ACS Style

Osborn, M.J.; Bhardwaj, A.; Bingea, S.P.; Knipping, F.; Feser, C.J.; Lees, C.J.; Collins, D.P.; Steer, C.J.; Blazar, B.R.; Tolar, J. CRISPR/Cas9-Based Lateral Flow and Fluorescence Diagnostics. Bioengineering 2021, 8, 23. https://doi.org/10.3390/bioengineering8020023

AMA Style

Osborn MJ, Bhardwaj A, Bingea SP, Knipping F, Feser CJ, Lees CJ, Collins DP, Steer CJ, Blazar BR, Tolar J. CRISPR/Cas9-Based Lateral Flow and Fluorescence Diagnostics. Bioengineering. 2021; 8(2):23. https://doi.org/10.3390/bioengineering8020023

Chicago/Turabian Style

Osborn, Mark J., Akshay Bhardwaj, Samuel P. Bingea, Friederike Knipping, Colby J. Feser, Christopher J. Lees, Daniel P. Collins, Clifford J. Steer, Bruce R. Blazar, and Jakub Tolar. 2021. "CRISPR/Cas9-Based Lateral Flow and Fluorescence Diagnostics" Bioengineering 8, no. 2: 23. https://doi.org/10.3390/bioengineering8020023

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