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Significance Herpes simplex virus-1 (HSV-1) is transmitted by contact between the infected tissues of a transmitter and those of an uninfected recipient. Effective spread requires that the lesions be small and not repulsive in appearance. For successful transmission HSV must control both its replication and its spread from infected to uninfected cells. Here we report that two HSV microRNAs (miRNAs) made late in productive infection are exported in exosomes. Ectopic expression in transfected cells before infection reduces the synthesis of viral gene products and the spread from infected to uninfected cells. The miRNAs accumulate in neurons after latent virus is induced to reactivate. The miRNAs conform to the expectations of gene products that control viral replication and spread.

miR-H28 and miR-H29 expressed late in productive infection are exported and restrict HSV-1 replication and spread in recipient cells