@article{17d46b54e4c84952a839bba9207c63b6,
title = "A Plasmid-Based Reverse Genetics System for Animal Double-Stranded RNA Viruses",
abstract = "Mammalian orthoreoviruses (reoviruses) are highly tractable experimental models for studies of double-stranded (ds) RNA virus replication and pathogenesis. Reoviruses infect respiratory and intestinal epithelium and disseminate systemically in newborn animals. Until now, a strategy to rescue infectious virus from cloned cDNA has not been available for any member of the Reoviridae family of dsRNA viruses. We report the generation of viable reovirus following plasmid transfection of murine L929 (L) cells using a strategy free of helper virus and independent of selection. We used the reovirus reverse genetics system to introduce mutations into viral capsid proteins σ1 and σ3 and to rescue a virus that expresses a green fluorescent protein (GFP) transgene, thus demonstrating the tractability of this technology. The plasmid-based reverse genetics approach described here can be exploited for studies of reovirus replication and pathogenesis and used to develop reovirus as a vaccine vector.",
keywords = "MICROBIO",
author = "Takeshi Kobayashi and Antar, {Annukka A.R.} and Boehme, {Karl W.} and Pranav Danthi and Eby, {Elizabeth A.} and Guglielmi, {Kristen M.} and Holm, {Geoffrey H.} and Johnson, {Elizabeth M.} and Maginnis, {Melissa S.} and Sam Naik and Skelton, {Wesley B.} and Wetzel, {J. Denise} and Wilson, {Gregory J.} and Chappell, {James D.} and Dermody, {Terence S.}",
note = "Funding Information: We thank Erik Barton, Craig Forrest, and Tim Peters for review of the manuscript and Dirk Reiter, Johannes Schilling, and Thilo Stehle for assistance in preparation of the figures. We thank Yoshihiro Kawaoka for plasmid p3E5EGFP and Tatsuo Miyamura and Koji Ishii for vaccinia virus rDIs-T7pol. This research was supported by a fellowship from the Naito Foundation (T.K.), Public Health Service awards T32 GM07347 (A.A.R.A. and E.A.E.), T32 CA09385 (K.W.B.), T32 GM08554 (K.M.G.), T32 AI49824 and F32 AI71440 (G.H.H.), T32 AI07611 (E.M.J.), T32 AI07281 (M.S.M.), K08 AI62862 (J.D.C.), R01 AI32539, and R37 AI38296, and the Elizabeth B. Lamb Center for Pediatric Research. Additional support was provided by Public Health Service awards P30 CA68485 for the Vanderbilt-Ingram Cancer Center and P60 DK20593 for the Vanderbilt Diabetes Research and Training Center. ",
year = "2007",
month = apr,
day = "19",
doi = "10.1016/j.chom.2007.03.003",
language = "English (US)",
volume = "1",
pages = "147--157",
journal = "Cell Host and Microbe",
issn = "1931-3128",
publisher = "Cell Press",
number = "2",
}