Following primary infection at the epidermis or mucosa, the human pathogen herpes simplex virus 1 (HSV1) establishes life-long latent infection in sensory neurons, reactivating periodically to cause cold sores, genital herpes or more severe clinical outcomes such as encephalitis or keratitis. Despite this clinical significance, no cure or licensed vaccine exists, potentially due to the difficulty of studying HSV1 in physiologically relevant conditions; although HSV1 infects complex stratified epithelia in the human, it has been primarily studied in monolayer cultures in the lab. Furthermore, HSV1-encoded accessory proteins that allow it to evade host responses may function differently in human skin compared to two-dimensional systems.   To address this, we have developed a 3D organotypic raft model using nTERT keratinocytes and have shown that these differentiate into a stratified structure resembling human skin. Infection of these rafts with fluorescently-tagged HSV1, either apically or basally to recapitulate primary infection from outside and reactivation from neurons respectively, demonstrated rapid spread through 3D cultures in just 3 days.   To investigate the requirement for the accessory protein UL13 protein kinase in skin infection, we have constructed a fluorescently-tagged UL13-deletion mutant (ΔUL13) which exhibits a greatly reduced plaque size in monolayer keratinocytes compared to Wt virus. While ΔUL13 also spread poorly in apically infected 3D cultures, it failed to spread at all during basal infection, indicating a key role for UL13 in HSV1 spread in human skin. These results demonstrate the importance of studying HSV1, and the role of HSV1 immune-evasion genes, in physiologically relevant 3D-skin models.