Use of short interfering RNAs (siRNAs) to investigate the role of specific tegument proteins in the intracellular movement of EHV-1 capsids at early times post-infection.

Upon entry into a cell, equine herpesvirus type 1 (EHV-1) capsids are actively transported to the nucleus where viral replication occurs. Previously, we showed that EHV-1 utilizes the microtubule network and the microtubule motor protein dynein to efficiently travel to the cell nucleus. While we identified some of the cellular components involved in capsid movement in the prior study, we did not identify any viral proteins that participate in this process. In the present study we investigate the use of siRNA technology to identify viral proteins that participate in the trafficking of viral capsids to the nucleus. We generated siRNA against green fluorescent protein (GFP) and showed that these GFP-specific siRNAs effectively reduced the expression of GFP produced from EHV-1. The data collected with the GFP siRNA suggest that we will be able to knockdown expression of specific virus genes and then assess whether this siRNA-mediated inhibition interferes with virus movement towards the nucleus. Current studies are being performed to assess whether one or more candidate tegument proteins, including EHV-1 ORF 40, are involved in virus trafficking to the nucleus.

Use of short interfering RNAs (siRNAs) to investigate the role of specific tegument proteins in the intracellular movement of EHV-1 capsids at early times post-infection. Presentation