A SIRNA SCREEN TO PROBE FOR HYDROXYLASES THAT CAN MODULATE THE REPLICATION OF DENGUE VIRUS

Abstract

Dengue virus (DENV) is the causative agent for dengue fever and the more severe dengue haemorrhagic fever / dengue shock syndrome which could result in death. Currently, with no effective vaccines or anti-virals available, 2.5 billion of the world¿s population is constantly at risk of DENV infection. In this study, a 384-well high-throughput RNAi-based screening platform was developed to screen genomic libraries for host factors that could modulate the replication of DENV in host cells. The application of the developed RNAi-based screening platform on a library of human protein hydroxylases established the association between hypoxia and the replication of DENV. Furthermore, this study has also shown for the first time, that replication of DENV2 could be modulated via the hypoxic pathway by hypoxic-inducible factors (HIFs). Finally, an expression study of the transcripts centered on the NF-kB pathway by PCR array revealed that the activation of the hypoxia-inducible pathway by HIFs resulted in an up-regulation of expression in the type 1 interferons (a and ß), which is likely to intervene in the replication of DENV2 in HuH-7 cells.