Author:
Blanchard Emmeline L.,Vanover Daryll,Bawage Swapnil Subhash,Tiwari Pooja Munnilal,Rotolo Laura,Beyersdorf Jared,Peck Hannah E.,Bruno Nicholas C,Hincapie Robert,Finn M.G.,Michel Frank,Lafontaine Eric R.,Hogan Robert J.,Zurla Chiara,Santangelo Philip J.
Abstract
ABSTRACTHere, 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.
Publisher
Cold Spring Harbor Laboratory
Cited by
4 articles.
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