- Ethan Hawke as Vincent Freeman, impersonating Jerome Eugene Morrow
- Mason Gamble as young Vincent
- Chad Christ as teenage Vincent
- Uma Thurman as Irene Cassini
- Jude Law as Jerome Eugene Morrow
- Loren Dean as Anton Freeman
- Vincent Nielson as young Anton
- William Lee Scott as teenage Anton
- Gore Vidal as Director Josef
- Xander Berkeley as Dr. Lamar
- Jayne Brook as Marie Freeman
- Elias Koteas as Antonio Freeman
- Maya Rudolph as delivery nurse
- Blair Underwood as geneticist
- Ernest Borgnine as Caesar
- Tony Shalhoub as German
- Alan Arkin as Detective Hugo
- Dean Norris as cop on the beat
- Ken Marino as sequencing technician
- Cynthia Martells as Cavendish
- Gabrielle Reece as Gattaca trainer
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