Generation of Infectious Recombinant Human Rotaviruses from Just 11 Cloned cDNAs Encoding the Rotavirus Genome
Generation of Infectious Recombinant Human Rotaviruses from Just 11 Cloned cDNAs Encoding the Rotavirus Genome
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DOI：
10.1128/jvi.02207-18
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发表时间：
2019-04-01
影响因子：
5.4
通讯作者：
Taniguchi, Koki
中科院分区：
文献类型：
--
作者：
Komoto, Satoshi;Fukuda, Saori;Taniguchi, Koki
完全从克隆的cDNA产生重组A组轮状病毒（RVA）仅描述了单个动物RVA株，猿猴SA 11-L2。我们最近开发了一个优化的RVA反向遗传学系统的基础上，只有RVA cDNA（11质粒系统），其中含有NSP 2和NSP 5基因的cDNA质粒的浓度是3-或5倍增加，相对于其他质粒。基于这种方法，我们使用11质粒系统产生了含有猴SA 11-L2病毒的VP 4基因的基于重组人RVA（HuRVA）的单克隆抗体病毒。除了这种单重组病毒外，还用11-质粒系统进行了一些修改，产生了真正的HuRVA（KU株）。我们的研究结果表明，11质粒系统涉及只是RVA cDNA可用于生产重组HuRVA和重组HuRVA为基础的reflectantvirus.IMPORTANCE人类A组轮状病毒（HuRVA）是一种主要的病原体，导致严重的腹泻，在世界各地的幼儿。在本文中，我们描述了重组HuRVA（株KU）的产生，从只有11个克隆的cDNA编码的HuRVA基因组的反向遗传学。重组HuRVA的生长特性与亲本RVA相似，为更好地理解HuRVA复制和发病机制提供了有力的工具。此外，操纵HuRVAs基因组“按顺序”的能力将有助于这种医学上重要的病毒的下一代疫苗生产和表达任何外源基因的临床载体的工程化。
The generation of recombinant group A rotaviruses (RVAs) entirely from cloned cDNAs has been described only for a single animal RVA strain, simian SA11-L2. We recently developed an optimized RVA reverse genetics system based on only RVA cDNAs (11-plasmid system), in which the concentration of cDNA plasmids containing the NSP2 and NSP5 genes is 3- or 5-fold increased in relation to that of the other plasmids. Based on this approach, we generated a recombinant human RVA (HuRVA)-based monoreassortant virus containing the VP4 gene of the simian SA11-L2 virus using the 11-plasmid system. In addition to this monoreassortant virus, authentic HuRVA (strain KU) was also generated with the 11-plasmid system with some modifications. Our results demonstrate that the 11-plasmid system involving just RVA cDNAs can be used for the generation of recombinant HuRVA and recombinant HuRVA-based reassortant viruses.IMPORTANCE Human group A rotavirus (HuRVA) is a leading pathogen causing severe diarrhea in young children worldwide. In this paper, we describe the generation of recombinant HuRVA (strain KU) from only 11 cloned cDNAs encoding the HuRVA genome by reverse genetics. The growth properties of the recombinant HuRVA were similar to those of the parental RVA, providing a powerful tool for better understanding of HuRVA replication and pathogenesis. Furthermore, the ability to manipulate the genome of HuRVAs "to order" will be useful for next-generation vaccine production for this medically important virus and for the engineering of clinical vectors expressing any foreign genes.