The director of the film was already quite familiar with the book by Che Guevara when the producers approached him to helm the film. He explains, "Ernesto Che Guevara's book had had a real impact on me because it is about a journey to discover not only one's identity and one's place in the world, but also about the search for what I think we could call a Latin American identity."

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To adapt Che Guevara's Motorcycle Diaries into a screenplay, Salles and the producers selected José Rivera. The screenwriter is an award-winning Puerto Rican-born playwright who had studied with Gabriel García Marquez at the Sundance Institute. Salles and Rivera conducted considerable research together, reading all the existing biographies on Ernesto Che Guevara. It would take Rivera two years of writing and rewriting, coupled with ongoing research, before the script was finished. To balance the film's portrayal of the two main characters, Rivera drew from both Che Guevara's travel diary and Granado's own account of the trip.

Rivera admits, "There were many challenges in writing this screenplay. How do you even begin to approach an iconic figure like Ernesto Che Guevara? I wanted to humanize the myth, yet also honor the memory and serve history. It was also important to give Alberto equal weight.The director found he was able to reconstruct the adventure and discover the present-day state of the places described in Ernesto Che Guevara's and Granado's accounts. Che Guevara's Motorcycle Diaries should and would be an Argentine film, all concerned firmly believed, so as to honor the origins of the film's main characters. Once the script was completed, the filmmakers began to prepare for filming. For the role of Ernesto Che Guevara, Mexican actor Gael García Bernal, whom the director describes as "one of the most talented actors of his generation," was offered the role. Drawn by the opportunity to incarnate a legend in his formative period, Bernal instantly accepted.