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Thirlby was born in New York City, the daughter of a construction worker. She was raised in Manhattan's East Village, attending school at Friends Seminary in the city's Gramercy neighborhood, where she graduated in a class of 57 students. She also attended French Woods Festival of the Performing Arts in upstate New York. She took classes at the American Globe Theatre, and briefly at the Royal Academy of Dramatic Art in London where she completed a stage combat course with the British Academy of Stage and Screen Combat (BASSC).

She plans to attend college once she has made a sufficient start to her acting career.

While still at high school, Thirlby was offered a role in The Secret. In 2006, she made her film debut in United 93 and her television debut in Kidnapped.

In the Sundance Audience Award-winning film The Wackness, a mid-1990s period piece, she plays Stephanie, a pothead from New York City. Thirlby stars opposite Josh Peck, who plays a drug dealer. Ben Kingsley stars as her stepfather. The film was released in the U.S. on July 3, 2008.

Thirlby was cast in the Judd Apatow-produced stoner comedy, Pineapple Express, as Seth Rogen's girlfriend, but was replaced by another actress after rehearsing for the film. She will however reuinite with Gordon Green on the animated TV pilot Good Vibes.

She made her stage debut in Farragut North, a play by Beau Willimon at the Atlantic Theater Company in New York City. The play is billed as "a classic tale of hubris set against a contemporary landscape — about the lust for power and the costs one will endure to achieve it." The cast included John Gallagher, Jr., Kate Blumberg and Isiah Whitlock, Jr., with Tony Award-winner Doug Hughes directing. The Off Broadway production ran from October 22, 2008 – November 29, 2008 with official opening on November 12.

In 2009, Thirlby will appear in a number of films including New York, I Love You, Margaret and Safety Glass and the HBO pilot Bored To Death.

She is attached to star in Parts Per Billion, Legion, Breaking Upwards which will reunite her with Darryl Wein, who directed her in Unlocked and Christmas In New York for Arlen Faber director John Hindman