- Rodrick Heffley as Gru
- Mr. Gerabit as Vector
- Mr. Jefferson as Dr. Nefario
- Holly Hills as Margo
- Ashley (from Alvin and the Chipmunks: The Road Chip) as Edith
- Lois Griffin (from Family Guy) as Agnes
- Anglerfish (from Finding Nemo) as Mr. Perkins
- Greg Heffley as Tim the Minion
- Rowley Jefferson as Phil the Minion
- Squrit (from Finding Nemo and Finding Dory) as Mark the Minion
- Bloat (from Finding Nemo and Finding Dory) as Kevin the Minion
- Gurgle (from Finding Nemo and Finding Dory) as Stuart the Minion
- Patty Farrell as Miss Hattie
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