Design and generation of a mutant gammaherpesvirus encoding shRNA from its native viral tRNA promoter

Gammaherpesviruses are associated with multiple types of tumour development and understanding the pathogenesis of these viruses has been the subject of many different studies. Throughout the lytic and latent life cycle, these viruses utilize numerous virally encoded miRNAs to regulate the key mechanisms of the infected cell in their favour. Therefore it is important to understand the miRNA and their mRNA target interactions for developing better therapeutics. In this study, the strategy and design of a short hairpin encoding RNA element targeting Blimp1 transcript in the context of viral infection is evaluated. This proof of principle experiment provides a mean to study important miRNA mRNA interactions in vivo. The very short promoter size of the Murine gammaherpesvirus 68 (MHV68) vtRNA4 and its ability to generate two shRNAs from a ~180 nucleotide sequence is useful if there is a size limit for the shRNA construct.