Dengue pp 285-299
Functional Genomics Approach for the Identification of Human Host Factors Supporting Dengue Viral Propagation
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 wordsRNA interference (RNAi) Dengue virus Yellow fever virus Whole-genome RNAi screening Whole-genome siRNA screening Dengue virus host factors Flavivirus
- 1.WHO’s Communicable Disease Global Atlas (2005) World Health Organization. www.who.int/denguenet. Accessed 9 Sept 2012.
- 2.Guzman MG, Halstead SB, Artsob H, Buchy P, Farrar J, Gubler DJ, Hunsperger E, Kroeger A, Margolis HS, Martinez E, Nathan MB, Pelegrino JL, Simmons C, Yoksan S, Peeling RW (2010) Dengue: a continuing global threat. Nat Rev Microbiol 8(Suppl 12):S7–S16. doi:10.1038/nrmicro2460 PubMedCrossRefGoogle Scholar
- 4.Krishnan MN, Ng A, Sukumaran B, Gilfoy FD, Uchil PD, Sultana H, Brass AL, Adametz R, Tsui M, Qian F, Montgomery RR, Lev S, Mason PW, Koski RA, Elledge SJ, Xavier RJ, Agaisse H, Fikrig E (2008) RNA interference screen for human genes associated with West Nile virus infection. Nature 455(7210):242–245. doi:10.1038/nature07207 PubMedCentralPubMedCrossRefGoogle Scholar