Reverse Genetics of Zika Virus.

We have established a reverse genetic system for Zika virus (ZIKV). Five shuttle plasmids were constructed and assembled into the full-length cDNA clone of ZIKV genome. To ensure the stability of the cDNA clone, we used a low copy vector (pACYC177) and a set of unique restriction enzyme sites on the ZIKV genome to assemble the full-length cDNA clone. A T7 promoter was engineered in front of the viral 5' UTR for in vitro transcription. A hepatitis delta virus ribozyme (HDVr) sequence was engineered following the viral 3' UTR for generation of the authentic 3' end of the RNA transcript.