Having acted on stage with her sister as a child, Gish was a prominent film star from 1912 into the 1920s, being particularly associated with the films of director D. W. Griffith. This included her leading role in the highest-grossing film of the silent era, Griffith's The Birth of a Nation (1915). Her other major films and performances from the silent era included Intolerance (1916), Broken Blossoms (1919), Way Down East (1920), Orphans of the Storm (1921), The White Sister (1923), La Bohème (1926), and The Wind (1928).
At the dawn of the sound era, she returned to the stage and appeared in film occasionally, with roles in the Western Duel in the Sun (1946) and the thriller The Night of the Hunter (1955). She was nominated for the Academy Award for Best Supporting Actress for Duel in the Sun. Gish also had major supporting roles in Portrait of Jennie (1948), A Wedding (1978), and Sweet Liberty (1986).