Crispr/cas9-based lateral flow and fluorescence diagnostics

Mark J. Osborn; Akshay Bhardwaj; Samuel P Bingea; Friederike Knipping; Colby J. Feser; Christopher J. Lees; Daniel P. Collins; Clifford J. Steer; Bruce R. Blazar; Jakub Tolar

doi:10.3390/BIOENGINEERING8020023

Crispr/cas9-based lateral flow and fluorescence diagnostics

Mark J. Osborn, Akshay Bhardwaj, Samuel P Bingea, Friederike Knipping, Colby J. Feser, Christopher J. Lees, Daniel P. Collins, Clifford J. Steer, Bruce R. Blazar, Jakub Tolar

Research output: Contribution to journal › Article › peer-review

23 Scopus citations

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/Cas9based 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.

Original language	English (US)
Article number	23
Pages (from-to)	1-20
Number of pages	20
Journal	Bioengineering
Volume	8
Issue number	2
DOIs	https://doi.org/10.3390/BIOENGINEERING8020023
State	Published - Feb 12 2021

Bibliographical note

Funding Information:
This research was funded by University of Minnesota Rapid COVID grants from the Medical School and Institute of Engineering in Medicine (M.J.O.). M.J.O. is also supported by a Saint Baldrick?s Foundation Scholar Award.

Funding Information:
Funding: This research was funded by University of Minnesota Rapid COVID grants from the Medical School and Institute of Engineering in Medicine (M.J.O.). M.J.O. is also supported by a Saint Baldrick’s Foundation Scholar Award.

Publisher Copyright:
© 2021 by the authors. Licensee MDPI, Basel, Switzerland.

Keywords

CRISPR/Cas9
Lateral flow assay
SARS-Co-V2

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10.3390/BIOENGINEERING8020023

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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/Cas9based 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.",

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Crispr/cas9-based lateral flow and fluorescence diagnostics

Abstract

Bibliographical note

Keywords

PubMed: MeSH publication types

UN SDGs

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