Establishment of Bi-segmented and Tri-segmented Reverse Genetics Systems to Generate Recombinant Pichinde Viruses

• Authors: Rekha Dhanwani, University of Minnesota; Qinfeng Huang, University of Minnesota; Shuiyun Lan, Emory University; Yanqing Zhou, University of Minnesota; Junkie Shao, University of Minnesota; Yuying Liang, Emory University; Hinh Ly, Emory University
• Journal Title: Methods in Molecular Biology
• Volume: Volume 1604
• Publisher: Humana Press | 2018-02-01, Pages 247-253
• Type of Work: Article | Post-print: After Peer Review
• Publisher DOI: http://doi.org/10.1007/978-1-4939-6981-4_19
• Subjects: Health Sciences, Pathology; Biology, Veterinary Science
• Keywords: arenavirus; reverse genetics; Pichindé virus

Abstract

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.