Dhanwani Rekha, Huang Qinfeng, Lan Shuiyun, Zhou Yanqing, Shao Junjie, Liang Yuying, Ly Hinh
Department of Veterinary and Biomedical Sciences, University of Minnesota, Twin Cities, 1988 Fitch Ave., 295 AS/VM Bldg, Saint Paul, MN, 55108, USA.
La Jolla Institute for Allergy and Immunology, La Jolla, CA, USA.
Methods Mol Biol. 2018;1604:247-253. doi: 10.1007/978-1-4939-6981-4_19.
Pichindé virus (PICV), isolated from rice rats in Colombia, South America, is an enveloped arenavirus with a bisegmented RNA genome. The large (L) genomic segment encodes the Z matrix protein and the L RNA-dependent RNA polymerase, whereas the small (S) genomic segment encodes the nucleoprotein (NP) and the glycoprotein (GPC). This article describes the successful development of reverse genetics systems to generate recombinant PICV with either a bisegmented or trisegmented genome. We have successfully demonstrated that these systems can generate high-titered and genetically stable replication-competent viruses from plasmid transfection into appropriate cell lines. These systems demonstrate the power and versatility of reverse genetic technology to generate recombinant arenaviruses for use in pathogenesis studies and as new viral vaccine vectors.
皮钦德病毒（PICV）于南美洲哥伦比亚的稻鼠体内分离得到，是一种具有包膜的沙粒病毒，其RNA基因组分为两段。大（L）基因组片段编码Z基质蛋白和L RNA依赖性RNA聚合酶，而小（S）基因组片段编码核蛋白（NP）和糖蛋白（GPC）。本文描述了反向遗传学系统的成功开发，该系统可用于产生具有双段或三段基因组的重组PICV。我们已成功证明，这些系统能够通过将质粒转染到合适的细胞系中，产生高滴度且遗传稳定的具有复制能力的病毒。这些系统证明了反向遗传技术在产生用于发病机制研究的重组沙粒病毒以及作为新型病毒疫苗载体方面的强大功能和多功能性。
