Treating Influenza and SARS-CoV-2 via mRNA-encoded Cas13a

Blanchard, E. L.; Vanover, D.; Bawage, S. S.; Tiwari, P. M.; Rotolo, L.; Beyersdorf, J.; Peck, H. E.; Bruno, N. C.; Hincapie, R.; Finn, M. G.; Michel, F.; Lafontaine, E. R.; Hogan, R. J.; Zurla, C.; Santangelo, P. J.

doi:10.1101/2020.04.24.060418

bioRxiv · 10.1101/2020.04.24.060418

Treating Influenza and SARS-CoV-2 via mRNA-encoded Cas13a

Blanchard, E. L.·Vanover, D.·Bawage, S. S.·Tiwari, P. M.·Rotolo, L.·Beyersdorf, J.·Peck, H. E.·Bruno, N. C.·Hincapie, R.·Finn, M. G.·Michel, F.·Lafontaine, E. R.·Hogan, R. J.·Zurla, C.·Santangelo, P. J.

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Abstract

Here, Cas13a has been used to target and mitigate influenza virus A (IAV) and SARS-CoV-2 using a synthetic mRNA-based platform. CRISPR RNAs (crRNA) against PB1 and highly conserved regions of PB2 were screened in conjunction with mRNA-encoded Cas13a. Screens were designed such that only guides that decreased influenza RNA levels in a Cas13-mediated fashion, were valid. Cas13a mRNA and validated guides, delivered post-infection, simulating treatment, were tested in combination and across multiplicities of infection. Their function was also characterized over time. Similar screens were performed for guides against SARS-CoV-2, yielding multiple guides that significantly impacted cytopathic effect. Last, the approach was utilized in vivo, demonstrating the ability to degrade influenza RNA in a mouse model of infection, using polymer-formulated, nebulizer-based mRNA delivery. Our findings demonstrate the applicability of Cas13a in mitigating respiratory infections both in vitro and in a mouse model, paving the way for future therapeutic use.

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Blanchard, E. L., Vanover, D., Bawage, S. S., Tiwari, P. M., Rotolo, L., Beyersdorf, J., Peck, H. E., Bruno, N. C., Hincapie, R., Finn, M. G., Michel, F., Lafontaine, E. R., Hogan, R. J., Zurla, C., Santangelo, P. J.. 2020-04-24. Treating Influenza and SARS-CoV-2 via mRNA-encoded Cas13a. https://doi.org/10.1101/2020.04.24.060418

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