Generation of fluorescent HCV pseudoparticles to study early viral entry events- involvement of Rab1a in HCV entry

Abstract

Understanding the early events in viral biology holds the key to the development of preventives. In this study fluorescent Hepatitis C Virus pseudoparticles have been generated where the envelope glycoprotein has a GFP tag. Using these pseudoparticles entry assays were conducted where the entry of the pseudoparticles was tracked via confocal microscopy. Using this system, fusion of host and viral membranes is predicted to occur within 15 minutes of entry in HCV. Using cells with a knockdown for Rab1a, HCV trafficking was observed to be altered, indicating a role of Rab1a in HCV trafficking. In conclusion, this study reports the generation and use of fluorescent pseudoparticles which may be used to understand the early events of viral entry. This system may be adapted for the study of other enveloped viruses as well. HighlightsO_LIFluorescent HCV pseudoparticles have been created to study early entry events. C_LIO_LIHCV entry tracking via confocal microscopy reveals fusion within 15 minutes. C_LIO_LIRab1a is important for HCV trafficking within the cell. C_LI