HCMV Latently infected Kasumi-3 transcription analysis.

HCMV latnecy within hematopoetic precursor cells was evaluated by infecting cells and establishing latency using a recombinant virus that allows for labeing of RNA species only in the context of a viral infection thereby allowing purification of these transcripts from mock infected cells. We used microarrays to detail the global programme of gene expression to identify changes in host transcription during HCMV latency. Kasumi-3 cells were either mock infected or infected with HCMV TB40e expressing T. gondii UPRT and IE2-2a-eGFP, Cells were sorted 96hpi to remove lytically replicating cells. Remaining cells were either uninfected or latenlty infected Kasumi-3 cells. This population was treated with 4-thoi uracil allowing incorporation of the label into the UPRT expressing cells only. After 6 hrs of labeling, RNA was isolated and used in a Biotin switch assay to purify labeled RNA from pre-existing RNA and RNA from uninfected cells. In parallel, mock infected cells were labeled with 4-thio Uradine adn used as a negative control. RNA transcription was monitored by affymetric array hybridization.