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 language||English (US)|
|Title of host publication||Methods in Molecular Biology|
|Publisher||Humana Press Inc.|
|Number of pages||7|
|State||Published - 2018|
|Name||Methods in Molecular Biology|
Bibliographical noteFunding 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.
- Pichindé virus
- Reverse genetics