1 High-throughput functional annotation of influenza A virus 1 genome at single-nucleotide resolution 2

17 A novel genome-wide genetics platform is presented in this study, which permits functional 18 interrogation of all point mutations across a viral genome in parallel. Here we generated the first 19 fitness profile of individual point mutations across the influenza virus genome. Critical residues 20 on the viral genome were systematically identified, which provided a collection of subdomain 21 data informative for structure-function studies and for effective rational drug and vaccine design. 22 Our data was consistent with known, well-characterized structural features. In addition, we 23 have achieved a validation rate of 68% for severely attenuated mutations and 94% for neutral 24 mutations. The approach described in this study is applicable to other viral or microbial genomes 25 where a means of genetic manipulation is available. 26 . CC-BY 4.0 International license peer-reviewed) is the author/funder. It is made available under a The copyright holder for this preprint (which was not . http://dx.doi.org/10.1101/005702 doi: bioRxiv preprint first posted online May. 31, 2014;