Development and application of dengue virus reverse genetic systems

The development of dengue virus "reverse genetic" systems based on full-length cDNA clones corresponding to the viral RNA genome has been an important technological platform for advancing dengue virus research. Mutations can be introduced into the genome to study their effect on virus replication and pathogenesis while attenuated or chimeric viruses can be constructed that are potential vaccine candidates. The deletion of the virus structural genes has led to the production of noninfectious, but replication competent viral subgenomes (termed replicons) that have been used to study viral replication and are useful for the screening of antiviral compounds. This article describes the development of dengue virus reverse genetic systems and protocols to manipulate the viral genome, recover infectious virus, and produce replicon-containing cell lines.