Construction of Genomic and Sub-Genomic Dengue Infectious Replicons

Methods Mol Biol. 2022:2409:77-96. doi: 10.1007/978-1-0716-1879-0_7.

Abstract

Dengue replicons are powerful tools used in studying virus biology as well as in high-throughput screening of drug candidates. Replicon constructs are developed as genomic (consists of all the viral protein genes) or sub-genomic (consists of only nonstructural protein genes) and are used to study various aspects of the virus life cycle such as genome replication and virus assembly. In addition, a replicon usually includes a reporter gene used in monitoring virus replication. In this chapter, we provide methods to develop both genomic and sub-genomic dengue replicons with a luciferase reporter and describe different assays to utilize these systems.

Keywords: Dengue replicon; Dengue virus serotype 2; Genomic replicon; Infectious dengue; Luciferase assay; Luciferase reporter; Plaque assay; Reporter virus; Sub-genomic replicon; Virus titer.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Dengue / genetics
  • Dengue Virus* / genetics
  • Genes, Reporter
  • Genomics
  • Humans
  • RNA, Viral
  • Replicon / genetics
  • Virus Replication / genetics

Substances

  • RNA, Viral