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Eliza Dushku was born in Boston, Massachusetts and came to the attention of casting agents when she was 10. She was chosen at the end of a five-month search throughout the United States for the lead role of Alice, opposite Juliette Lewis in the film That Night. In 1993, Dushku landed the role of Pearl alongside Robert De Niro and Leonardo DiCaprio in This Boy's Life, a role that opened many doors. The following year, she played the teenage daughter of Arnold Schwarzenegger and Jamie Lee Curtis in True Lies. She also had parts as the daughter of Paul Reiser's Donny in Bye Bye Love.

Dushku then took on the role of Faith Lehane, a Slayer much more troubled than the main character Buffy Summers in TV's Buffy the Vampire Slayer. Though initially planned as a five-episode role, the character became so popular that she stayed on for the entirety of the third season and returned for a two-part appearance in season four, after which the remainder of her original story arc was played out as part of the first season of the Buffy spinoff series Angel.

In 2000, Dushku starred in Soul Survivors, reuniting her with Race The Sun co-star Casey Affleck. She followed that up with the cheerleader comedy Bring It On with Kirsten Dunst, which was a surprising success at the box office that spawned straight-to-DVD sequels. In 2001, she appeared in The New Guy opposite DJ Qualls and City by the Sea opposite Robert De Niro and James Franco. The latter film garnered attention from a wider adult audience and very good reviews. The same year, Kevin Smith invited

Dushku to be a part of Jay and Silent Bob Strike Back, where Dushku co-starred with Shannon Elizabeth, Ali Larter and Ben Affleck.

In 2003, she appeared in Wrong Turn, a horror film in which Dushku had the starring role; and The Kiss, an independent comedy-drama. Starting that same year, she also starred in a new Fox TV series, Tru Calling, where she played the main character.

Dushku starred in an off-Broadway production entitled Dog Sees God from December 2005, playing Van's sister, a character paralleled with Lucy from the original Peanuts comic strip that the play production is based on.

Most recently, she can be seen in Miller/Savin's Bottle Shock and the upcoming On Broadway (filmed in her native Boston) She has also signed a development deal with Fox Broadcasting and 20th Century Fox to produce and star in a new TV series. Under the deal, she is teaming with Joss Whedon of Buffy fame to create a new series entitled Dollhouse, premiering on Fox TV in January.