Dujardin first became famous on the French talent show Graines de star in 1996 as part of the comedy group Nous C Nous, formed by members of the Carré blanc theater. From 1999 to 2003 he starred in the French version of the comedy television series Un gars, une fille before transitioning into a film career. In 2005 he starred as the titular character in the popular comedy film Brice de Nice and performed the soundtrack of the film, "Le Casse de Brice".
In 2006 Dujardin portrayed the character Hubert Bonisseur de La Bath, agent OSS 117, in OSS 117: Cairo, Nest of Spies in a role which earned him a César Award nomination for Best Actor, something of exceptional rarity for a performance in comedy. In 2007, directed by Jan Kounen, he scored in 99F (99 francs), an existential parody of an advertising mule. In 2009 he again played OSS 117 in the film OSS 117: Lost in Rio.
He received the award for best actor at the Cannes Film Festival in 2011.
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