Lynne Marie Stewart, who plays Jimmy, the snake farm owner, was Miss Yvonne in the original Pee-wee's Playhouse (1986) TV series.
Along with wearing makeup and using tape on his face, visual effects were used to "de-age" Paul Reubens's face. The Pee-wee films and the television series were used as references.
The female bank robbers are patterned after Varla, Rosie and Billie from Russ Meyer's cult classic Faster, Pussycat! Kill! Kill! (1965).
This is the first Pee-wee Herman film in 28 years.