Directed genetic modification of African horse sickness virus by reverse genetics

  • Elaine Vermaak Department of Microbiology and Plant Pathology, University of Pretoria, Pretoria, South Africa
  • Duncan J. Paterson Sir William Dunn School of Pathology, University of Oxford, Oxford, United Kingdom
  • Andele Conradie Department of Microbiology and Plant Pathology, University of Pretoria, Pretoria, South Africa
  • Jacques Theron Department of Microbiology and Plant Pathology, University of Pretoria, Pretoria, South Africa
Keywords: orbivirus, core transcripts, genome modification, vaccine

Abstract

African horse sickness virus (AHSV), a member of theĀ OrbivirusĀ genus in the family Reoviridae, is an arthropod-transmitted pathogen that causes a devastating disease in horses with a mortality rate greater than 90%. Fundamental research on AHSV and the development of safe, efficacious vaccines could benefit greatly from an uncomplicated genetic modification method to generate recombinant AHSV. We demonstrate that infectious AHSV can be recovered by transfection of permissive mammalian cells with transcripts derived in vitro from purified AHSV core particles. These findings were expanded to establish a genetic modification system for AHSV that is based on transfection of the cells with a mixture of purified core transcripts and a synthetic T7 transcript. This approach was applied successfully to recover a directed cross-serotype reassortant AHSV and to introduce a marker sequence into the viral genome. The ability to manipulate the AHSV genome and engineer specific mutants will increase understanding of AHSV replication and pathogenicity, as well as provide a tool for generating designer vaccine strains.

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Published
2015-07-27