title: Generation of a variety of stable Influenza A reporter viruses by genetic engineering of the NS gene segment.
abstract: Influenza A viruses (IAV) pose a constant threat to the human population and therefore a better understanding of their fundamental biology and identification of novel therapeutics is of upmost importance. Various reporter-encoding IAV were generated to achieve these goals, however, one recurring difficulty was the genetic instability especially of larger reporter genes. We employed the viral NS segment coding for the non-structural protein 1 (NS1) and nuclear export protein (NEP) for stable expression of diverse reporter proteins. This was achieved by converting the NS segment into a single open reading frame (ORF) coding for NS1, the respective reporter and NEP. To allow expression of individual proteins, the reporter genes were flanked by two porcine Teschovirus-1 2A peptide (PTV-1 2A)-coding sequences. The resulting viruses encoding luciferases, fluorescent proteins or a Cre recombinase are characterized by a high genetic stability in vitro and in mice and can be readily employed for antiviral compound screenings, visualization of infected cells or cells that survived acute infection.
authors.affiliation: 1] Institute for Virology, University of Freiburg, 79104 Freiburg, Germany [2] Faculty of Biology, University of Freiburg, 79104 Freiburg, Germany.
authors.forename: Peter
authors.initials: P
authors.lastname: Reuther
authors.affiliation: Institute for Virology, University of Freiburg, 79104 Freiburg, Germany.
authors.forename: Kristina
authors.initials: K
authors.lastname: Göpfert
authors.affiliation: 1] Institute for Virology, University of Freiburg, 79104 Freiburg, Germany [2] Faculty of Biology, University of Freiburg, 79104 Freiburg, Germany [3].
authors.forename: Alexandra H
authors.initials: AH
authors.lastname: Dudek
authors.forename: Monika
authors.initials: M
authors.lastname: Heiner
authors.forename: Susanne
authors.initials: S
authors.lastname: Herold
authors.affiliation: Institute for Virology, University of Freiburg, 79104 Freiburg, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Schwemmle
mesh_terms: D000818:Animals; D004285:Dogs; D017930:Genes, Reporter; D057809:HEK293 Cells; D006801:Humans; D009980:Influenza A virus / Q000235:genetics / Q000378:metabolism; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D061985:Madin Darby Canine Kidney Cells; D051379:Mice; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: C085820:INS1 protein, influenza virus; D017361:Viral Nonstructural Proteins; D008156:Luciferases