Only one herpes simplex virus type 1 (HSV-1) gene is expressed in sensory neurons of latently infected animals and humans, yielding two RNAs, called latency-associated transcripts (LATs). The LATs appear to modulate virus reactivation. In mice and rabbits the 5' origins, kinetics of synthesis, and splicing pattern of the LATs are well established. Because these details of LAT structure and expression have not been defined in humans, we sought to do so. Using primer extension and Northern hybridization analyses, we demonstrate that in human trigeminal ganglia, the smaller (1.35 kb) HSV-1 transcript differs from the larger (1.85 kb) LAT by excision of an intron near its 5' end; they are otherwise colinear, and 5' coterminal. In infected cells only the 1.85 kb LAT is detected. Its expression is inhibited by cycloheximide or acyclovir, indicating this LAT is synthesized late in the viral replicative cycle. All of these features of the LATs in humans are consistent with those reported in rabbits and mice and further validate the animal models of human HSV-1 infection.