While in college, Rai did a few modelling assignments. Following appearances in several television commercials, she entered the Miss India pageant, in which she placed second. She was then crowned Miss World 1994, after which she began receiving offers to act in film. She made her acting debut with the 1997 Tamil film Iruvar and had her first Hindi film release in Aur Pyaar Ho Gaya that same year. Her first commercial success was the Tamil romantic drama Jeans (1998), following which she achieved wider success and two Best Actress awards at Filmfare for her performances in Hum Dil De Chuke Sanam (1999) and Devdas (2002). She garnered critical appreciation for portraying Rabindranath Tagore’s heroine, Binodini, in the Bengali film Chokher Bali (2003), a depressed woman in the relationship drama Raincoat (2004) and as Kiranjit Ahluwalia in the British drama film Provoked (2006). Rai’s greatest commercial successes have been the adventure film Dhoom 2 (2006), the historical romance Jodhaa Akbar (2008), and the science fiction film Enthiran (2010). She also received praise for portraying a nurse in the drama Guzaarish (2010). Rai took a sabbatical from film acting in 2010, and made her comeback with the thriller Jazbaa (2015).
Rai married actor Abhishek Bachchan in 2007 with whom she has a daughter. Her off-screen roles include duties as a brand ambassador for several charity organisations and campaigns. She is a Goodwill Ambassador for the Joint United Nations Programme on AIDS (UNAIDS). In 2003, she was the first Indian actress to be a jury member at the Cannes Film Festival.