Transposon mutagenesis of the Zika virus genome highlights regions essential for RNA replication and restricted for immune evasion

RNA-seq of Zika virus libraries To investigate Zika virus (ZIKV) genetic flexibility, we used transposon mutagenesis to add 15-nucleotide insertions throughout the MR766 ZIKV genome and deep sequenced the viable genomes. The ZIKV transposon library was constructed and rescued as described before: Heaton, N.S., Sachs, D., Chen,C.J., Hai, R., and Palese, P. (2013). Genome-wide mutagenesis of influenza virus reveals unique plasticity of the hemagglutinin and NS1 proteins. PNAS. 110, 20248-20253, and Fulton, B.O., Sachs, D., Beaty, S.M., Won, S.T., Lee, B., Palese, P., and Heaton, N.S. (2015). Mutational Analysis of Measles Virus Suggests Constraints on Antigenic Variation of the Glycoproteins. Cell Rep. 1(9): 11331–1338.