Having run across an ad for an open casting call and attending her first audition, Rodriguez beat 350 other applicants to win her first role in the low-budget 2000 independent film, Girlfight. With her performance as Diana Guzman, a troubled teen who decides to channel her aggression by training to become a boxer, Rodriguez accumulated several awards and nominations for the role in independent circles, including major acting accolades from the National Board of Review, Deauville Film Festival, Independent Spirit Awards, Gotham Awards, Las Vegas Film Critics Sierra Awards, and many others. The film itself took home a top prize at the Sundance and won Award of the Youth at the Cannes Film Festival.
Subsequently, she has had notable roles in other successful movies, including The Fast and the Furious, Resident Evil, Blue Crush, and S.W.A.T. In 2004, Rodriguez lent her voice to the video game Halo 2, playing a Marine. She also provided the voice of Liz Ricarro in the Cartoon Network series IGPX. From 2005 to 2006, she played tough cop Ana Lucia Cortez on the television series Lost during the show's second season (the character's first appearance was a flashback on season 1's finale, Exodus: Part 1), and returned for a cameo in the second episode of the show's fifth season The Lie in 2009 and again in the penultimate episode of the series What They Died For in 2010. In 2006, Rodriguez was featured in her own episode of G4's show Icons.
In 2008, she appeared in Battle in Seattle alongside Charlize Theron. Rodriguez next appeared in the fourth installment of the The Fast and the Furious franchise, which was titled simply Fast & Furious and released to theaters on April 3, 2009. Rodriguez is in James Cameron's high-budget sci-fi adventure film Avatar, which was released to theaters on December 18, 2009. She also recently wrapped filming on Trópico de Sangre, an independent film based on the Dominican Republic's historic Mirabal sisters who were assassinated in 1960 by the Dominican dictator Trujillo for opposing his rule.
As of 20 July 2010, Rodriguez's films have grossed $1,272,734,719 in the United States and $3,686,521,043 worldwide.
In 2010, Rodriguez appeared in Robert Rodriguez's Machete, and will next star alongside Aaron Eckhart in the sci-fi film Battle: Los Angeles.