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Born and raised in Sacramento, California, Chastain developed an interest in acting from an early age. In 1998, she made her professional stage debut as Shakespeare's Juliet. After studying acting at the Juilliard School, she was signed to a talent holding deal with the television producer John Wells. She was a recurring guest star in several television series, including Law & Order: Trial by Jury. She also took on roles in the stage productions of Anton Chekhov's play The Cherry Orchard in 2004 and Oscar Wilde's tragedy Salome in 2006.

Chastain made her film debut in the drama Jolene (2008), and gained wide recognition for her starring roles in the dramas Take Shelter (2011) and The Tree of Life (2011). Her performance as an aspiring socialite in The Help (2011) earned her a nomination for the Academy Award for Best Supporting Actress. In 2012, she won a Golden Globe Award and received a nomination for the Academy Award for Best Actress for playing a CIA analyst in the thriller Zero Dark Thirty. Chastain made her Broadway debut in a revival of The Heiress in the same year. Her highest-grossing releases came with the science fiction films Interstellar (2014) and The Martian (2015), and the horror film It Chapter Two (2019), and she continued to receive critical acclaim for her performances in the dramas A Most Violent Year (2014), Miss Sloane (2016), Molly's Game (2017) and The Good Nurse (2022). For her portrayal of Tammy Faye in the biopic The Eyes of Tammy Faye (2021), which she also produced, Chastain won the Academy Award for Best Actress.

In television, Chastain starred in drama miniseries Scenes from a Marriage (2021) and George & Tammy (2022). For the latter, she won a SAG Award. Her performance also garnered her nominations for a Primetime Emmy Award and a Golden Globe Award.

Chastain is the founder of the production company Freckle Films, which was created to promote diversity in film. She is vocal about mental health issues, as well as gender and racial equality. She is married to fashion executive Gian Luca Passi de Preposulo, with whom she has two children.