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Cook's latest voice-over role is for the video game Yakuza in which she voices the role of Reina.
Cook first gained national attention in 1997, when she was featured in the famous This Is Your Brain on Drugs PSA television advertisement, in which she proceeds to destroy a kitchen with a frying pan as she lists the important things in life that heroin harms.
In 2011, she was selected by the Obama administration as a Champion of Change for Arts Education.
In 2017, Rachael Leigh Cook reprised her "This Is Your Brain on Drugs" role twenty years later for a PSA by the Drug Policy Alliance critiquing the War on Drugs and its contribution to mass incarceration, structural racism and poverty.
She starred in the independent film Broken Kingdom, which was directed by her husband Daniel Gillies.
She also appeared in a Funny or Die sketch with Chad Michael Murray.
In 2008, she guest-starred in an episode of the USA Network series Psych as Abigail Lytar. In February 2010, Cook signed on to play the female lead role in Fox TV's comedy pilot Nirvana.
In 2012, Cook signed on to play the female lead role in the TNT crime drama series Perception opposite Eric Mc Cormack.
She made her debut as an actress in the 1995 film The Baby-Sitters Club.
She took the lead role in 2001's Josie and the Pussycats, which turned out to be a box office failure.