Dushku was born in Watertown, Massachusetts, the daughter of Philip Richard George Dushku, an administrator and teacher in the Boston Public Schools, and Judith Ann "Judy" Dushku (née Rasmussen), a political science professor.
Dushku came to the attention of casting agents when she was 10.
She was chosen in a five-month search for the lead role of Alice in the film That Night.
In 1993, Dushku landed a role as Pearl alongside Robert De Niro and Leonardo Di Caprio in This Boy's Life, a role that she said opened a lot of doors.
She starred in Open Graves, a 2008 horror-thriller about a satanic game co-starring Mike Vogel.In an interview, Dushku talked about how Dollhouse and her reconnection with Whedon came about: I invited Joss Whedon to lunch after I did the business deal with Fox.We'd had a cool relationship in the past and I so wanted to do something else, and I wanted to get back into a television show." – and there's some guy sat on the sofa with a bottle of beer and a moustache, and a big gut. Dushku turned down a role in a spin-off of Buffy The Vampire Slayer which would have been about Faith.She has had many roles as a "bad girl" in movies and relishes the opportunities.This was the second Fox pilot in which she was cast, but not broadcast.On October 1, 2005, she announced at Wizard World Boston that shooting had begun for Nobel Son in which she would star with Alan Rickman, Danny De Vito and Bill Pullman.In an interview with Maxim in May 2001, Dushku says of her roles, "It's easy to play a bad girl: You just do everything you've been told not to do, and you don't have to deal with the consequences, because it's only acting." Dushku starred in an Off-Broadway production entitled Dog Sees God from December 2005, playing "Van's sister", a character paralleled with Lucy Van Pelt from the Peanuts comic strip on which the play production is based.She quit in February 2006 along with other members of the cast amidst rumors of abuse from the producer, which were latery-drama for Fox.but her agent asked her to submit a videotape audition for a show starring another of his clients, Sarah Michelle Gellar.After reading the script, Dushku rushed to a local Claire's to purchase dark makeup and other appropriate accessories for the part, Faith on Buffy the Vampire Slayer.