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The Pixar and Disney animation studios' chief creative officer, John Lasseter, confirmed the casting at a presentation in Los Angeles, Reuters reports.
Winfrey will play the character of Eudora, the mother of the main character Princess Tiana (Anika Noni Rose).
The film, scheduled for release in late 2009, is set in the French Quarter of New Orleans and Randy Newman has written six new songs for its soundtrack.
Winfrey has been involved in several animated and fantasy films in the past, most recently playing Judge Bumbleton in Bee Movie.
More: Oprah Winfrey, Movies
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At the Movies
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Movies Interviews
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Box Office Charts
'The Woman in Black' wins UK box officeThe Woman in Black beats The Muppets and Star Wars at the the UK box office.
