Treatment of influenza and SARS-CoV-2 infections via mRNA-encoded Cas13a in rodents
Treatment of influenza and SARS-CoV-2 infections via mRNA-encoded Cas13a in rodents
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DOI：
10.1038/s41587-021-00822-w
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发表时间：
2021-02-03
影响因子：
46.9
通讯作者：
Santangelo, Philip J.
中科院分区：
文献类型：
--
作者：
Blanchard, Emmeline L.;Vanover, Daryll;Santangelo, Philip J.
Cas 13 a被递送到小鼠和仓鼠的肺中抑制流感病毒和SARS-CoV-2.Cas13a的复制已被用于在细胞培养中靶向RNA病毒，但尚未在动物模型中证实其功效。在这项研究中，我们使用信使RNA（mRNA）编码的Cas 13 a来减轻流感病毒A和严重急性呼吸综合征冠状病毒2（SARS-CoV-2）感染小鼠和仓鼠，分别。我们设计了针对流感病毒PB 1和PB 2高度保守区，以及针对SARS-CoV-2复制酶和核衣壳基因的CRISPR RNA（crRNA），并筛选了在细胞培养中最有效降低病毒RNA水平的crRNA。我们使用喷雾器将聚合物配制的Cas 13 a mRNA和经验证的指导物递送至呼吸道。在小鼠中，Cas 13 a在感染后递送时有效降解肺组织中的流感RNA，而在仓鼠中，Cas 13 a递送减少了SARS-CoV-2复制并减轻了症状。我们的研究结果表明，Cas 13 a介导的致病病毒靶向可以减轻呼吸道感染。
Cas13a delivered to the lung of mice and hamsters inhibits replication of influenza virus and SARS-CoV-2.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.