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Open AccessArticle

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

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Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

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