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Significance Human cytomegalovirus (HCMV), a herpesvirus that persists latently in much of the world’s population, can cause serious disease in immune-compromised individuals and is a leading cause of birth defects. Despite its importance, however, our understanding of HCMV latency remains enigmatic. This is due in part to a technical bottleneck in which virus mutants can be generated only in a specific cell type. To address this limitation, here we describe a technology that enables cell-specific silencing of a desired viral transcript. This allows generation of the virus without impacting its biology during acute infection while retaining the capacity to study specific gene function in the context of a desired cell type.

miRNA-mediated targeting of human cytomegalovirus reveals biological host and viral targets of IE2