engleski

Transcriptomic profile of murine cytomegalovirus in vitro and in vivo

Human cytomegalovirus (HCMV), a ubiquitous beta-herpesvirus, is capable of establishing lifelong persistence with minimal or no damage to its host. However, in immunosuppressed and immunologically immature patients it can cause grave disease and even death. Additionally, HCMV infection is the leading cause of organ transplant failure and birth defects. Unfortunately pathogenesis of HCMV infection is still poorly understood while better treatments and effective vaccine is needed. Murine cytomegalovirus (MCMV) is biologically similar and genetically related to HCMV and is therefore considered to be an excellent model for in vivo studies. To date, many new immunoevasins have been characterized thanks to a plethora of deletion mutants. However, currently used genomic map of the MCMV relies heavily on ORFs predicted using bioinformatics tools and verified by genomic tilling arrays. Such analyses can miss certain ORFs, rare alternative splicing sites and do not give information about the exact 5’ and 3’ ends of the transcripts. Therefore, the main goal of this project was to characterize all of the viral transcription products that accumulate in infected cells. Similar transcriptomic analysis in HCMV revealed many novel genes and gene products, and our analysis of MCMV’s transcriptome, reveals unexpected complexity of the MCMV genome which could not have been envisioned with ORF prediction software alone. We have detected a number of new transcripts and novel splice-variants, along with several sense-antisense pairs. Currently we’re employing deep sequencing to analyze small RNAs which may have been missed in the original library, as well as viral and mouse transcription profiles in vivo in different organs in the course of infection. Interestingly, many of the most abundantly expressed viral genes, including the top five, have no known function. We have also detected significant transcription from regions previously annotated as noncoding. Taken together, these data stress out the importance of transcriptomic analysis in construction of genomic maps. Our study is among the first to investigate the transcriptome of virus and host both in the course of the infection. Analysis of host responses in different organs has a tremendeous potential to identify new biomarkers and provide new insights into global and organ specific immune responses to this virus.

transcriptome murine cmv mcmv rnaseq

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