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Generation of recombinant rotaviruses from just 11 cDNAs encoding a viral genome.
Virus Research ( IF 2.5 ) Pub Date : 2020-06-24 , DOI: 10.1016/j.virusres.2020.198075
Satoshi Komoto 1 , Saori Fukuda 1 , Riona Hatazawa 1 , Takayuki Murata 1 , Koki Taniguchi 1
Affiliation  

Reverse genetics technology allows one to engineer replication-competent viruses from cloned cDNAs at will. Since the establishment of the initial reverse genetics system for species A rotaviruses (RVAs) requiring a helper virus in 2006, attempts have been successfully made to improve this technology. Efficient generation of replication-competent RVAs is now possible from just 11 T7-driven plasmids encoding an RVA genome when the quantity ratio of the two rescue T7-driven plasmids for the NSP2 and NSP5 segments is increased by 3-fold in relation to that of the other nine plasmids (11 plasmid-only system). Further, it is now possible to generate recombinant RVAs even with severely less efficient infectivity by using the 11 plasmid-only system, which has not been possible with the existing approaches. More importantly, the 11 plasmid-only system does not need any helper expression plasmid, and thus this simplest and robust system has a clear advantage over the existing systems in terms of safety. This 11 plasmid-only system should contribute to the development of safe next-generation vaccines and vaccine vectors.



中文翻译:

仅从编码病毒基因组的11个cDNA生成重组轮状病毒。

逆向遗传学技术使人们可以从克隆的cDNA中随意工程改造具有复制能力的病毒。自2006年建立了需要辅助病毒的A类轮状病毒(RVA)的初始反向遗传学系统以来,已成功进行了改进该技术的尝试。现在,当NSP2和NSP5区段的两个抢救性T7驱动质粒的数量比相对于NSP2的3倍增加时,仅由11个编码RVA基因组的T7驱动质粒即可有效生成具有复制能力的RVA。其他9个质粒(仅11个质粒系统)。此外,现在可以通过使用仅11个质粒的系统来产生重组RVA,即使其感染效率大大降低,这是现有方法无法实现的。更重要的是,仅11个质粒的系统不需要任何辅助表达质粒,因此,在安全性方面,这种最简单,最可靠的系统明显优于现有系统。仅11个质粒的系统应有助于开发安全的下一代疫苗和疫苗载体。

更新日期:2020-06-26
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