Lee Chang-Won
Food Animal Health Research Program, Ohio Agricultural Research and Development Center, The Ohio State University, 1680 Madison Avenue, Wooster, OH, 44691-4096, USA,
Methods Mol Biol. 2014;1161:37-50. doi: 10.1007/978-1-4939-0758-8_4.
Reverse genetics is the creation of a virus from a full-length cDNA copy of the viral genome, referred to as an "infectious clone," and is one of the most powerful genetic tools in modern virology. Since its development in 1999, plasmid-based reverse genetics has been effectively applied to numerous aspects of influenza studies which include revolutionizing the production of seasonal and pandemic influenza vaccine seed strains. Although continual improvement in reverse genetics system is being made in different laboratories for the efficient rescue of the influenza virus, the basic concept of synthesizing viral RNA using RNA polymerase I remains the same. Coupled with in vitro mutagenesis, reverse genetics can be applied widely to accelerate progress in understanding the influenza virus life cycle, the generation of customized vaccine seed strains, development of live-attenuated vaccines, and the use of influenza virus as vaccine and gene delivery vectors.
反向遗传学是通过病毒基因组的全长cDNA拷贝构建病毒，这种cDNA拷贝被称为“感染性克隆”，它是现代病毒学中最强大的遗传工具之一。自1999年开发以来，基于质粒的反向遗传学已有效地应用于流感研究的诸多方面，其中包括彻底改变季节性和大流行性流感疫苗种子株的生产。尽管不同实验室在不断改进反向遗传学系统以高效拯救流感病毒，但利用RNA聚合酶I合成病毒RNA的基本概念保持不变。结合体外诱变，反向遗传学可广泛应用于加速对流感病毒生命周期的理解、定制疫苗种子株的产生、减毒活疫苗的开发以及将流感病毒用作疫苗和基因递送载体等方面的研究进展。
