Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York 10029, USA.
Global Health and Emerging Pathogens Institute, Division of Infectious Diseases, Icahn School of Medicine at Mount Sinai, New York, NY 10029.
Nat Biotechnol. 2013 Sep;31(9):844-847. doi: 10.1038/nbt.2666. Epub 2013 Aug 11.
Recent gain-of-function studies in influenza A virus H5N1 strains revealed that as few as three-amino-acid changes in the hemagglutinin protein confer the capacity for viral transmission between ferrets. As transmission between ferrets is considered a surrogate indicator of transmissibility between humans, these studies raised concerns about the risks of gain-of-function influenza A virus research. Here we present an approach to strengthen the biosafety of gain-of-function influenza experiments. We exploit species-specific endogenous small RNAs to restrict influenza A virus tropism. In particular, we found that the microRNA miR-192 was expressed in primary human respiratory tract epithelial cells as well as in mouse lungs but absent from the ferret respiratory tract. Incorporation of miR-192 target sites into influenza A virus did not prevent influenza replication and transmissibility in ferrets, but did attenuate influenza pathogenicity in mice. This molecular biocontainment approach should be applicable beyond influenza A virus to minimize the risk of experiments involving other pathogenic viruses.
最近的甲型流感病毒 H5N1 株的功能获得性研究表明，血凝素蛋白中仅三个氨基酸的变化即可使病毒在雪貂之间传播。由于雪貂之间的传播被认为是人与人之间传播能力的替代指标，这些研究引起了人们对甲型流感病毒功能获得性研究风险的担忧。在这里，我们提出了一种加强功能获得性流感实验生物安全性的方法。我们利用物种特异性内源性小 RNA 来限制流感病毒的嗜性。具体而言，我们发现 microRNA miR-192 在人呼吸道上皮细胞以及小鼠肺部中表达，但在雪貂呼吸道中不存在。将 miR-192 靶位点整合到甲型流感病毒中并不会阻止流感在雪貂中的复制和传播，但会减轻流感在小鼠中的致病性。这种分子生物控制方法应该适用于除甲型流感病毒以外的其他致病病毒，以最大限度地降低涉及其他致病病毒的实验的风险。
