MHV-68 RNA-Seq for PolyA Study
RNA-Seq analysis and polyadenylation site detection in Murine Gammaherpesvirus 68
TBD
Recent tiled array studies have provided high resolution data on the transcriptomes of human and mouse gammaherpesviruses including the identification of a number of regions of unexpected RNA expression.  While these microarray-based studies have provided a unique view of viral RNA expression, they are limited in their ability to define individual RNA transcripts.  We have utilized directional RNA-Seq to generate strand-specific information on the transcriptome of murine gammaherpesvirus 68 in late-stage lytic infection.  These results confirmed the existence of the previously identified regions of RNA expression.  In addition, we have developed software that enabled us to mine RNA-Seq data for the 3’ end of transcripts.  We identified over 200 putative polyadenylation sites across the viral genome.  Finally, we developed a novel validation method that combines traditional 3’ RACE with next generation sequencing to test thirteen of our predicted polyadenylation sites.