After gaining wide attention in a Parker Pen commercial with Amitabh Bachchan, D'Souza began her acting career with Tujhe Meri Kasam in 2003.

D'Souza received her first Filmfare Award in 2006 for her performance in the Telugu romantic film, Bommarillu.

In 2008, Genelia gave critically acclaimed performances in Santosh Subramaniam, a Tamil remake of Bommarillu, and the Bollywood movie Jaane Tu... Ya Jaane Na.

Genelia is the brand ambassador of Fanta, Virgin Mobile India, Fastrack, LG Mobiles, Garnier Light, Margo, and Perk in India.

Genelia won the Golden Nandi award in 2006 for her performance in Telugu romantic film Bommarillu.

Genelia D'Souza was one of the judges at the grand finale of Gladrags Mega Model and Manhunt 2009 contest on 28 March 2009.

D'Souza is set to appear in It's My Life, which would be a remake of Bomarillu in Hindi with Harman Baweja.