PT  - JOURNAL ARTICLE
AU  - Emmeline L. Blanchard
AU  - Daryll Vanover
AU  - Swapnil Subhash Bawage
AU  - Pooja Munnilal Tiwari
AU  - Laura Rotolo
AU  - Jared Beyersdorf
AU  - Hannah E. Peck
AU  - Nicholas C Bruno
AU  - Robert Hincapie
AU  - M.G. Finn
AU  - Frank Michel
AU  - Eric R. Lafontaine
AU  - Robert J. Hogan
AU  - Chiara Zurla
AU  - Philip J. Santangelo
TI  - Treating Influenza and SARS-CoV-2 via mRNA-encoded Cas13a
AID  - 10.1101/2020.04.24.060418
DP  - 2020 Jan 01
TA  - bioRxiv
PG  - 2020.04.24.060418
4099  - http://biorxiv.org/content/early/2020/04/24/2020.04.24.060418.short
4100  - http://biorxiv.org/content/early/2020/04/24/2020.04.24.060418.full
AB  - 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.Competing Interest StatementThe authors have declared no competing interest.