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Gene therapy using adenoviruses has shown tremendous promise in animal and human models in the past. The tumor responses achieved by these viruses were either because of their oncolytic properties or therapeutic genes expressed from the genome of the virus. We have previously shown enhanced viral replication in cell models that have lost or lower expression of the cyclin dependent kinases inhibitor (p21/Waf-1). Beside the early onset of many viral genes in the p21/Waf-1 knock out cell model, we observed a significantly higher copy number for the viral VA1 but non VA2 non-coding RNA transcripts. In this study we investigated the effect of adenoviral VA1 non-coding RNA on the landscape of cellular proteins.