When the Duke of Milan is brutally murdered, the Assassin Giovanni Auditore is dispatched to investigate. What he uncovers implicates Italy's most powerful families reaching all the way back to the Vatican itself. As Giovanni draws closer to the truth, he becomes hunted himself. He must expose the conspirators before he joins their ever-growing list of victims.
| Actor | Role |
|---|---|
 Romano OrzariGiovanni Auditore | Giovanni Auditore |
 Manuel TadrosRodrigo Borgia | Rodrigo Borgia |
 Jesse RathFederico Auditore | Federico Auditore |
 Devon BostickEzio Auditore | Ezio Auditore |
 Alex IvanoviciLorenzo Medici | Lorenzo Medici |
 Michel PerronUberto Alberti | Uberto Alberti |
| AArthur GrosserPope Sixtus IV | Pope Sixtus IV |
 Peter MillerGalleazzo Maria Sforza | Galleazzo Maria Sforza |
 Harry StandjofskiSilvio Barbarigo | Silvio Barbarigo |
 Frank FontaineShawn Baichoo | Shawn Baichoo |
 Claudia FerriMaria Auditore | Maria Auditore |
 Devery JacobsClaudia Auditore | Claudia Auditore |
 Patrick KertonLampugnari | Lampugnari |
 Marcello BezinaMonk/Servant | Monk/Servant |
 Bianca LeBlancEzio's Girlfriend | Ezio's Girlfriend |
 Roc LaFortunePrisoner | Prisoner |