- Dr. Frank N. Furter as Mario
- Janet Weiss as Pauline
- Brad Majors as Luigi
- Riff Raff as Toad
- Magenta as Princess Daisy
- Columbia as Princess Peach
- Dr. Everett V. Scott as Bowser
- Rocky Horror as Donkey Kong
- Eddie as Yoshi
- The Ciminologist as Wario
- Ralph Hapschatt as Waluigi
- Betty Munroe Hapschatt as Princess Rosalina
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