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

• Type of Material: Article
• 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
• Language: English
• Date: 2018-02-01
• Publisher: Humana Press
• Publication Version: Author Accepted Manuscript (After Peer Review)
• Copyright Statement: © Springer Science+Business Media LLC 2018
• Final Published Version (URL): http://dx.doi.org/10.1007/978-1-4939-6981-4_19
• Title of Journal or Parent Work: Methods in Molecular Biology
• ISSN: 1064-3745
• Volume: 1604
• Start Page: 247
• End Page: 253
• Grant/Funding Information: This work was supported in part by the NIAID/NIH through the new-direction awards mechanism of the SERCEB grant (U54-AI057157) to YL and HL; by the NIAID/NIH R01 AI083409 to YL; and R01 AI093580 and R56 AI091805 to HL.
• Keywords: reverse genetics; arenavirus; Pichindé virus
• Research Categories: Health Sciences, Pathology; Biology, Veterinary Science

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.