Treatment of influenza and SARS-CoV-2 infections via mRNA-encoded Cas13a in rodents

Authors

• Emmeline L. Blanchard, Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA, USA.
• Daryll Vanover, Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA, USA.
• Swapnil Subhash Bawage, Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA, USA.
• Pooja Munnilal Tiwari, Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA, USA.
• Laura Rotolo, Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA, USA.
• Jared Beyersdorf, Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA, USA.
• Hannah E. Peck, Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA, USA.
• Nicholas C. Bruno, School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, GA, USA.
• Robert Hincapie, School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, GA, USA.
• Frank Michel, Department of Veterinary Biosciences and Diagnostic Imaging, College of Veterinary Medicine University of Georgia, Athens, GA, USA.
• Jackelyn Murray, Department of Infectious Diseases, College of Veterinary Medicine University of Georgia, Athens, GA, USA.
• Heena Sadhwani, Department of Biology, Georgia State University, Atlanta, GA, USA.
• Bob Vanderheyden, Kennesaw State University
• M G. Finn, School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, GA, USA.
• Margo A. Brinton, Department of Biology, Georgia State University, Atlanta, GA, USA.
• Eric R. Lafontaine, Department of Infectious Diseases, College of Veterinary Medicine University of Georgia, Athens, GA, USA.
• Robert J. Hogan, Department of Veterinary Biosciences and Diagnostic Imaging, College of Veterinary Medicine University of Georgia, Athens, GA, USA.
• Chiara Zurla, Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA, USA. chiara@gatech.edu.
• Philip J. Santangelo, Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA, USA. philip.santangelo@bme.gatech.edu.

Department

School of Data Science and Analytics

Document Type

Article

Publication Date

6-1-2021

Abstract

Cas13a has been used to target RNA viruses in cell culture, but efficacy has not been demonstrated in animal models. In this study, we used messenger RNA (mRNA)-encoded Cas13a for mitigating influenza virus A and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection in mice and hamsters, respectively. We designed CRISPR RNAs (crRNAs) specific for PB1 and highly conserved regions of PB2 of influenza virus, and against the replicase and nucleocapsid genes of SARS-CoV-2, and selected the crRNAs that reduced viral RNA levels most efficiently in cell culture. We delivered polymer-formulated Cas13a mRNA and the validated guides to the respiratory tract using a nebulizer. In mice, Cas13a degraded influenza RNA in lung tissue efficiently when delivered after infection, whereas in hamsters, Cas13a delivery reduced SARS-CoV-2 replication and reduced symptoms. Our findings suggest that Cas13a-mediated targeting of pathogenic viruses can mitigate respiratory infections.

Journal Title

Nature biotechnology

Volume

39

Issue

6

First Page

717

Last Page

726

Digital Object Identifier (DOI)

10.1038/s41587-021-00822-w