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Establishment of a Chimeric, Replication-Deficient Influenza A Virus Vector by Modulation of Splicing Efficiency

2010; American Society for Microbiology; Volume: 85; Issue: 5 Linguagem: Inglês

10.1128/jvi.01650-10

1098-5514

Markus Wolschek, Elisabeth Samm, Helena Seper, Sanda Šturlan, И. Л. Кузнецова, Cornelia Schwager, Alexandra Khassidov, Christian Kittel, Thomas Muster, Andrej Egorov, Michael Bergmann,

Viral Infections and Immunology Research

Segment 8 of the influenza A virus codes for two proteins (NS1 and NS2/NEP) via splicing. Here, we developed a viral vector expressing a cytokine or chemokine instead of the interferon antagonist NS1. To achieve both the desired genetic stability and high transgene expression levels, NS2/NEP mRNA splicing efficacy had to be fine-tuned by modification of splicing elements. Expression levels of secreted foreign proteins could be further enhanced by fusing the N-terminal 13 amino acids of NS1 with an IgK-derived secretion signal peptide. Thus, the first start codon was used for translation initiation of both NS2/NEP and the foreign protein.