Establishment of bisegmented and trisegmented reverse genetics systems to generate recombinant pichindé viruses

Rekha Dhanwani, Qinfeng Huang, Shuiyun Lan, Yanqing Zhou, Junjie Shao, Yuying Liang, Hinh Ly

Research output: Chapter in Book/Report/Conference proceedingChapter

4 Scopus citations

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.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages247-253
Number of pages7
DOIs
StatePublished - 2018

Publication series

NameMethods in Molecular Biology
Volume1604
ISSN (Print)1064-3745

Bibliographical note

Funding Information:
We thank Prof. Dr. K. Conzelmann (Ludwig-Maximilians-Universität, Germany) for providing the BSRT7-5 cells, and K. Curtis (USAMRIID, USA) for providing the pUC19-HDVT7t vector and Lassa genomic information. This work was supported in part by the NIAID/NIH through the new direction awards program of the SERCEB grant (U54-AI057157) to Y.L. and H.L., by the NIAID/NIH R01 AI083409 to Y.L., and R01 AI093580 and R56 AI091805 to H.L.

Keywords

  • Arenavirus
  • Pichindé virus
  • Reverse genetics

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