- Aleksandr Vladimirovich Serebryakov (Александр Владимирович Серебряков) – a retired university professor, who has lived for years in the city on the earnings of his late first wife's rural estate, managed for him by Vanya and Sonya.
- Helena Andreyevna Serebryakov (Yelena) (Елена Андреевна Серебрякова) – Professor Serebryakov's young and beautiful second wife. She is 27 years old.
- Sofia Alexandrovna Serebryakov (Sonya) (Софья Александровна Серебрякова) – Professor Serebryakov's daughter from his first marriage. She is of a marriageable age but is considered plain.
- Maria Vasilyevna Voynitsky (Мария Васильевна Войницкая) – the widow of a privy councilor and mother of Vanya (and of Vanya's late sister, the Professor's first wife).
- Ivan Petrovich Voynitsky ("Uncle Vanya") (Иван Петрович Войницкий) – Maria's son and Sonya's uncle, the title character of the play. He is 47 years old.
- Mikhail Lvovich Astrov (Михаил Львович Астров) – a middle aged country doctor.
- Ilya Ilych Telegin (Илья Ильич Телегин; nicknamed "Waffles" for his pockmarked skin) – an impoverished landowner, who now lives on the estate as a dependent of the family.
- Marina Timofeevna (Марина Тимофеевна) – an old nurse.
- A Workman
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