Genome-Wide RNAi Screen for Viral Replication in Mammalian Cell Culture
Influenza infections are considered a global threat to public health and cause seasonal epidemics and recurring pandemics. High mutation rates facilitate the generation of viral escape mutants rendering vaccines and drugs directed against virus-encoded targets ineffective. One alternative approach that could prevent viral escape is the targeting of host cell determinants that are temporarily dispensable for the host but crucial for virus replication. Here, we report a genome-wide RNAi screening approach in mammalian cell culture system that led us to the identification of several host cell genes influencing influenza A virus replication. Interestingly, the majority of the identified host gene products are indispensable for viral replication of a broad range of influenza viruses ranging from the highly pathogenic avian H5N1 strain to the current pandemic swine-origin H1N1 strain. Our results provide a new approach to explore virus–host interactions and to identify promising antiviral targets.
Key wordsInfluenza A virus H1N1 H5N1 RNAi siRNA
The authors would like to thank N. Machuy, Y. Shin, K.-P. Pleissner, A. Mäurer, and T. Wolff for significant scientific input in setting up the screen, M. Drabkina, G. Heins, D. Khalil, and D. Manntz for technical support. We also thank A. Pekosz for providing the influenza luciferase reporter construct and S. Becker for the pandemic H1N1 influenza strain. This work was supported through the EU FP6 project RIGHT (LSHB-CT-2004-005276), ERA-Net Pathogenomics (grant no. 0313938A) and a grant from FCI to T.F.M., and the RiNA network Berlin to T.R. and T.F.M.
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