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 journalArticlepeer-review

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 languageEnglish (US)
Article number23
Pages (from-to)1-20
Number of pages20
JournalBioengineering
Volume8
Issue number2
DOIs
StatePublished - Feb 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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