Protocol

Dengue

Volume 1138 of the series Methods in Molecular Biology pp 285-299

Date:

Functional Genomics Approach for the Identification of Human Host Factors Supporting Dengue Viral Propagation

  • Nicholas J. BarrowsAffiliated withProgram in Cell and Molecular Biology, Department of Molecular Genetics and Microbiology, Center for RNA Biology, Duke University Medical Center
  • , Sharon F. JamisonAffiliated withDepartment of Molecular Genetics and Microbiology, Center for RNA Biology, Duke University Medical Center
  • , Shelton S. BradrickAffiliated withDepartment of Molecular Genetics and Microbiology, Center for RNA Biology, Duke University Medical Center
  • , Caroline Le SommerAffiliated withDepartment of Molecular Genetics and Microbiology, Center for RNA Biology, Duke University Medical Center
  • , So Young KimAffiliated withDepartment of Molecular Genetics and Microbiology, Center for RNA Biology, Duke University Medical Center
  • , James PearsonAffiliated withDepartment of Molecular Genetics and Microbiology, Center for RNA Biology, Duke University Medical Center
  • , Mariano A. Garcia-BlancoAffiliated withDepartment of Molecular Genetics and Microbiology, Center for RNA Biology, Duke University Medical Center

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Abstract

Dengue virus (DENV) is endemic throughout tropical regions of the world and there are no approved treatments or anti-transmission agents currently available. Consequently, there exists an enormous unmet need to treat the human diseases caused by DENV and block viral transmission by the mosquito vector. RNAi screening represents an efficient method to expand the pool of known host factors that could become viable targets for treatments or provide rationale to consider available drugs as anti-DENV treatments. We developed a high-throughput siRNA-based screening protocol that can identify human DENV host factors. The protocol herein describes the materials and the procedures necessary to screen a human cell line in order to identify genes which are either necessary for or restrict DENV propagation at any stage in the viral life cycle.

Key words

RNA interference (RNAi) Dengue virus Yellow fever virus Whole-genome RNAi screening Whole-genome siRNA screening Dengue virus host factors Flavivirus