A Plasmid-Based Reverse Genetics System for Animal Double-Stranded RNA Viruses

@article{citeulike:2620851, abstract = {Summary 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 [sigma]1 and [sigma]3 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.}, author = {Kobayashi, Takeshi and Antar, Annukka A. and Boehme, Karl W. and Danthi, Pranav and Eby, Elizabeth A. and Guglielmi, Kristen M. and Holm, Geoffrey H. and Johnson, Elizabeth M. and Maginnis, Melissa S. and Naik, Sam and Skelton, Wesley B. and Wetzel, Denise J. and Wilson, Gregory J. and Chappell, James D. and Dermody, Terence S. }, citeulike-article-id = {2620851}, doi = {http://dx.doi.org/10.1016/j.chom.2007.03.003}, journal = {Cell Host \& Microbe}, keywords = {molecular\_methods, other\_reovirus}, month = {April}, number = {2}, pages = {147--157}, posted-at = {2008-04-01 20:32:42}, priority = {2}, title = {A Plasmid-Based Reverse Genetics System for Animal Double-Stranded RNA Viruses}, url = {http://dx.doi.org/10.1016/j.chom.2007.03.003}, volume = {1}, year = {2007} }

TY - JOUR ID - citeulike:2620851 L3 - citeulike-article-id:2620851 TI - A Plasmid-Based Reverse Genetics System for Animal Double-Stranded RNA Viruses JF - Cell Host & Microbe VL - 1 IS - 2 SP - 147 EP - 157 N2 - Summary 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 [sigma]1 and [sigma]3 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. KW - molecular_methods KW - other_reovirus AU - Kobayashi, Takeshi AU - Antar, Annukka AU - Boehme, Karl AU - Danthi, Pranav AU - Eby, Elizabeth AU - Guglielmi, Kristen AU - Holm, Geoffrey AU - Johnson, Elizabeth AU - Maginnis, Melissa AU - Naik, Sam AU - Skelton, Wesley AU - Wetzel, Denise AU - Wilson, Gregory AU - Chappell, James AU - Dermody, Terence PY - 2007/04/19/ UR - http://dx.doi.org/10.1016/j.chom.2007.03.003 ER -