Abstract:

Gammaherpes viruses are associated with the development of cancers in various types and the pathogenesis of these viruses has been the subject of many studies. These viruses code a large number of viral-based microRNAs (miRNAs) to regulate the key mechanisms of the infected cell with two different life cycles called lithium and latent. Therefore, it is important to understand myRNA and their mRNA target interactions to develop better therapeutic substances. Here, we showed with a lusiferaz experiment that a shRNA, produced by viral tRNA and targeted the Blimp1 transcript, can significantly reduce the expression of the target gen in the viral infection stage from a statistical point of view. This experimental design aims to provide a concept proof in terms of testing the important miRNA-mRNA interactions. Murine Gammaherpesvirus 68 virus tRNA4, which has a fairly short promotor size, can produce two shRNAs from a series of about 180 nucleotid lengths. If there is only a limited area for the description of the desired shRNA, the viral tRNA promoter offers an effective shRNA description system.

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Abstract:

Gammaherpesviruses are associated with multiple types of tumor development and understanding the pathogenesis of these viruses has been the subject of various studies. Throughout the lytic and latent life cycle, these viruses utilize numerous virally encoded microRNAs (miRNAs) to regulate the key mechanisms of the infected cell in their favor. 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 recombinant virus expressing a short hairpin RNA (shRNA) element targeting the host B-lymphocyte-induced maturation protein 1 (Blimp1) transcript was evaluated. Here we have shown that viral tRNA-driven expression of anti-Blimp1 shRNA is able to reduce the target gene expression at a statistically significant level as assessed by luciferase assay during virus infection. This proof-of-principle experiment provides a means to study important miRNA-mRNA interactions in vivo. Further, the very short promoter of the murine gammaherpesvirus 68 (MHV68) viral tRNA (vtRNA4) has the ability to generate two shRNAs from a ~180 nucleotide sequence. If there is a size limit for the shRNA construct, viral tRNA promoter provides an effective shRNA expression system.