RNA-seq transcriptome analysis of HCMV (AD169) and mock infected MRC5 fibroblasts
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ABSTRACT: We employed RNA-seq to map the transcriptome of human MRC5 fibroblasts during HCMV infection with AD169. These data will highlight the ways in which the HCMV infection alters RNA levels during infection.
Project description:We employed RNA-seq to map the transcriptome of human MRC5 fibroblasts during HCMV infection with AD169 and AD169-ΔUL26 strains. These data will highlight the ways in which the HCMV UL26 protein alters host gene transcripton during infection.
Project description:Infection of human fibroblasts with either human cytomegalovirus strain AD169 or a recombinant virus deleted for HCMV US17 Illumina HT-12 beadarrays were used to quantitate levels of host transcripts at either 12 or 96 hpi.
Project description:We report ChIP-seq analsyis of human fibroblasts infected with HCMV strains AD169 and TB40E. ChIP was performed at 20 hpi for IE2 and 3 dpi for IE2 and UL84.
Project description:Infection of human fibroblasts with either human cytomegalovirus strain AD169 or a recombinant virus deleted for HCMV US17 Illumina HT-12 beadarrays were used to quantitate levels of host transcripts at either 12 or 96 hpi. Cells were infected in biological triplicates at both 12 and 96 hours post infetion
Project description:Human cytomegalovirus (HCMV) is a widespread virus and can establish life-long latent infection in large populations. In order to establish persistent and latent infection in healthy individuals, HCMV encodes a large array of proteins that can modulate different components and pathways of host cells. It has been reported that pUL138 encoded by UL133-UL138 polycistronic locus promotes a latent infection in primary CD34+ HPCs infected in vitro. In this study, a recombinant HCMV, namely HanUL138del, was constructed by deleting the UL138 locus of Han, a clinic HCMV strain. Then a comparative quantitative proteomic analysis of Han and HanUL138del infected MRC5 cells was performed, aiming to study the effect of pUL138 on host cell in the context of HCMV infection. Our result indicated that at the early phase of HCMV infection, innate immune response was differentially activated, while at late phase of HCMV infection, multiple host proteins were differentially expressed, between Han or HanUL138del infected cells.
Project description:To investigate how HCMV alters monocyte gene expression over time, we collected RNA from HCMV-infected primary human monocytes (3 donors) from 4 hours to 6 weeks post infection and analyzed the differential gene expression by RNAseq.
Project description:Human cytomegalovirus (HCMV) is a significant cause of disease in immune-compromised adults and immune naïve newborns. No vaccine exists to prevent HCMV infection, and current antiviral therapies have toxic side effects that limit the duration and intensity of their use. There is thus an urgent need for new strategies to treat HCMV and repurposing existing drugs as antivirals is an attractive approach. Virus-induced changes in infected cells are often driven by changes in cellular kinase activity, leading us to hypothesize that defining the complement of kinases (the kinome), whose activity or expression is altered during infection would identify existing kinase inhibitors that could be repurposed as new antivirals. To this end, we applied a recently developed technique, MIB-MS kinome profiling, to quantitatively measure perturbations in >240 cellular kinases simultaneously in cells infected with a laboratory-adapted (AD169) or clinical (TB40E) HCMV strain using a label-free approach. Significant time-dependent changes for multiple kinases including cell cycle, receptor tyrosine and mitotic kinases were observed. Based on these data, we tested antiviral activity of 14 kinase inhibitors, and the most potent inhibitor blocked HCMV early gene expression and viral DNA accumulation. These results show the utility of kinome profiling to screen kinase inhibitors that can be repurposed as antivirals.