Japanese journal of infectious diseases

Development of a novel Dengue-1 virus replicon system expressing secretory Gaussia luciferase for analysis of viral replication and discovery of antiviral drugs.

PMID 24858611


Replicon systems have been used for high-throughput screening of anti-dengue virus (anti-DENV) inhibitors and for understanding mechanisms of viral replication. In the present study, we constructed novel DENV-1 replicons encoding Gaussia luciferase that was secreted into the culture medium. Two types of constructs were generated: RNA-based and DNA-based. Each type was translated in an internal ribosome entry site (IRES)-dependent or IRES-independent manner. Among these constructs, the DNA-based replicon employing IRES-dependent translation (DGL2) produced the highest titer. Luciferase levels in the culture medium revealed that the DGL2 replicon was inhibited by ribavirin (a well-known DENV inhibitor) at levels similar to those measured for drug inhibition of multi-round DENV-1 infection. These results indicate that the DNA-based IRES-driven DENV-1 replicon may facilitate studies on viral replication and antiviral compound discovery.