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J. Virol. doi:10.1128/JVI.01290-07
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Discrete clusters of viral encoded miRNAs are associated with complementary strands of the genome and the 7.2- kb stable intron in murine cytomegalovirus

Division of Pathway Medicine, University of Edinburgh, Edinburgh, UK.; Foundation for Research and Technology, Hellas, FORTH, Greece

^* To whom correspondence should be addressed. Email: a.buck{at}ed.ac.uk.

	Abstract

The prevalence and importance of microRNAs (miRNAs) in viral infection is increasingly relevant. Eleven miRNAs were previously identified in human cytomegalovirus (HCMV), however, miRNA content in murine CMV (MCMV), which serves as an important in vivo model for CMV infection, has not previously been examined. We have cloned and characterized 17 novel miRNAs that originate from at least 12 precursor miRNAs in MCMV and are non homologous to HCMV miRNAs. In parallel, we applied a computational analysis, using a support vector machine approach, to identify potential precursor miRNAs in MCMV. Four of the top 10 predicted precursor sequences were cloned in this study and the combination of computational and cloning analysis demonstrates that MCMV has the capacity to encode miRNAs clustered throughout the genome. On the basis of drug sensitivity experiments for resolving kinetic class of expression, we show that the MCMV miRNAs are both early and late gene products. Notably, the MCMV miRNAs occur on complementary strands of the genome in specific regions, a feature which has not previously been observed for viral miRNAs. One cluster of miRNAs occurs in close proximity to the 5' splice site of the previously identified 7.2 kb stable intron, implying a variety of potential regulatory mechanisms for MCMV miRNAs.

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