Gena Rowlands was born in Madison, Wisconsin and was raised in Cambria, Wisconsin. Her father, Edwin Myrwyn Rowlands, was a banker and a state legislator and her mother, Mary Allen, was a painter and housewife originally from Arkansas. The family moved to Washington, D.C. in 1939, when Edwin was appointed to a position in the U.S. Department of Agriculture, then to Milwaukee, Wisconsin in 1942, when he was appointed as branch manager of the Office of Price Administration, and later to Minneapolis, Minnesota. Gena attended the University of Wisconsin from 1947 to 1950, where she was a popular student already renowned for her beauty. She left for New York City to study drama at the American Academy of Dramatic Arts.

Gena performed in stock companies, 1951–1953, and at the Provincetown Playhouse. In 1952–1955, she went from understudy to lead role in the original Broadway production of The Seven Year Itch, and toured in a national production of the play. She opened and starred in Middle of the Night (1956) opposite film icon Edward G. Robinson.

She guest starred in several anthology television series, including Robert Montgomery Presents, Kraft Television Theatre and Studio One, among many others, in 1955. She made her film debut in The High Cost of Loving in 1958. In 1961-1962, she starred as the deaf-mute wife of Robert Lansing on NBC's 87th Precinct television series. In 1964, she was cast in ABC's Peyton Place nighttime soap opera.

Teaming with her husband, writer and director John Cassavetes, whom she married in 1954, Rowlands starred in the television production Staccato, and the films A Child Is Waiting, Faces, Gloria (nomination for Academy Award for Best Actress), Love Streams, Minnie and Moskowitz, She's So Lovely, and A Woman Under the Influence (Academy Award nomination for Best Actress).

In 1985, she played the mother in the critically acclaimed made-for-TV movie An Early Frost. In recent years, she has appeared in Paulie and in Mira Nair's HBO movie, Hysterical Blindness for which she won her third Emmy.

She was recently seen in The Notebook, which was directed by her son, Nick Cassavetes, and co-starred James Garner Ryan Goslingand Rachel McAdams. In 2004 she won her first Daytime Emmy for her role as Mrs. Evelyn Ritchie in The Incredible Mrs. Ritchie. To name a few, Gena Rowlands has been nominated for: two Academy Awards; six Emmy nominations, and one Daytime Emmy; eight Golden Globes; three Satellite Awards; and one SAG Award. Some of her notable wins include: a Silver Berlin Bear; three Emmy Awards and one Daytime Emmy; two Golden Globes; two National Board of Review Awards; two Satellite Awards; and one Prize San Sebastián.

In 2005, she appeared opposite Kate Hudson Peter Sarsgaard and John Hurt in the gothic thriller The Skeleton Key.