- Don Pedro, Prince of Aragon
- Benedick, of Padua; a lord, companion of Don Pedro
- Claudio, of Florence; a lord, companion of Don Pedro
- Balthasar, attendant on Don Pedro, a singer
- Don John, "the Bastard Prince," brother of Don Pedro and the main villain
- Borachio and Conrade, followers of Don John
- Leonato, governor of Messina
- Hero, Leonato's daughter
- Beatrice, an orphan, Leonato's niece
- Antonio, an old man, brother of Leonato
| - Margaret, waiting-gentlewoman attendant on Hero
- Ursula, waiting-gentlewoman attendant on Hero
- Friar Francis, a priest
- Dogberry, the grand constable in charge of Messina's night watch
- Verges, the Headborough, Dogberry’s partner
- A Sexton, the judge of the trial of Borachio
- The Watch,watchmen of Messina
- A Boy, serving Benedick
- Attendants and messengers
- Innogen, a ghost character included in early editions as Leonato's wife
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