Conradie, Andele M.Stassen, LieselHuismans, H. (Henk), 1942-Potgieter, Christiaan A.Theron, Jacques2016-10-192016-12Conradie, AM, Stassen, L, Huismans, H, Potgieter, CA & Theron, J 2016, 'Establishment of different plasmid only-based reverse genetics systems for the recovery of African horse sickness virus', Virology, vol. 499, pp. 144-155.0042-6822 (print)1096-0341 (online)10.1016/j.virol.2016.07.010http://hdl.handle.net/2263/57362In an effort to simplify and expand the utility of African horse sickness virus (AHSV) reverse genetics, different plasmid-based reverse genetics systems were developed. Plasmids containing cDNAs corresponding to each of the full-length double-stranded RNA genome segments of AHSV-4 under control of a T7 RNA polymerase promoter were co-transfected in cells expressing T7 RNA polymerase, and infectious AHSV-4 was recovered. This reverse genetics system was improved by reducing the required plasmids from 10 to five and resulted in enhanced virus recovery. Subsequently, a T7 RNA polymerase expression cassette was incorporated into one of the AHSV-4 rescue plasmids. This modified 5-plasmid set enabled virus recovery in BSR or L929 cells, thus offering the possibility to generate AHSV-4 in any cell line. Moreover, mutant and cross-serotype reassortant viruses were recovered. These plasmid DNA-based reverse genetics systems thus offer new possibilities for investigating AHSV biology and development of designer AHSV vaccine strainsen© 2016 Elsevier Inc. All rights reserved. Notice : this is the author’s version of a work that was accepted for publication in Virology. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. A definitive version was subsequently published in Virology, vol. 499, pp. 144-155, 2016. doi : 10.1016/j.virol.2016.07.010.OrbivirusReassortmentReoviridaeReverse geneticsT7 RNA polymeraseVirus rescuedsRNAAfrican horse sickness virus (AHSV)Postprint Article