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.
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