title: Development of a novel Dengue-1 virus replicon system expressing secretory Gaussia luciferase for analysis of viral replication and discovery of antiviral drugs.
abstract: 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.
authors.affiliation: Laboratory of Primate Model, Experimental Research Center for Infectious Diseases, Institute for Virus Research, Kyoto University.
authors.forename: Fumihiro
authors.initials: F
authors.lastname: Kato
authors.forename: Takeshi
authors.initials: T
authors.lastname: Kobayashi
authors.forename: Shigeru
authors.initials: S
authors.lastname: Tajima
authors.forename: Tomohiko
authors.initials: T
authors.lastname: Takasaki
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Miura
authors.forename: Tatsuhiko
authors.initials: T
authors.lastname: Igarashi
authors.forename: Takayuki
authors.initials: T
authors.lastname: Hishiki
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D002460:Cell Line; D006224:Cricetinae; D003715:Dengue / Q000188:drug therapy / Q000821:virology; D003716:Dengue Virus / Q000187:drug effects / Q000235:genetics*; D055808:Drug Discovery / Q000379:methods*; D017930:Genes, Reporter; D008156:Luciferases / Q000235:genetics* / Q000378:metabolism; D012367:RNA, Viral / Q000235:genetics; D012093:Replicon / Q000235:genetics*; D012254:Ribavirin / Q000494:pharmacology; D014776:Virus Cultivation; D014779:Virus Replication / Q000187:drug effects* / Q000235:genetics
chemical_list: D000998:Antiviral Agents; D012367:RNA, Viral; D012254:Ribavirin; D008156:Luciferases