After completing her education, Jentsch pursued a career in acting. She enrolled at the renowned Ernst Busch Academy of Dramatic Arts in Berlin, laying the foundation for her future success. Her early years in the theater saw her join the Munich Kammerspiele ensemble, where she showcased her exceptional talent by taking on iconic roles such as Desdemona in Shakespeare's "Othello" and Sophocles' "Antigone". Her impressive performances did not go unnoticed, as she received the Max Reinhardt Prize in 2000 for her work in the Berlin production of "The Persians". The magazine "Theater aktuell" later named her the best young actress in 2002, a testament to her remarkable stage presence.
In addition to her thriving theater career, Jentsch made her cinematic debut in Judith Kennel's successful film "Angry Kisses" in 2000. Her television appearances included episodes of "Praxis Bülowbogen" (2000),"And the Bride knew of nothing" (2002),and "Tatort" (2003). Her filmography continued to grow with roles in "Julietta" (2001) by Christoph Stark, "My Brother, the Vampire" (2001) by Sven Taddicken, and "Downfall" (2004) under the direction of Oliver Hirschbiegel. Jentsch then took on the lead role in Hans Weingartner's "The Fat Years Are Over" (2004),earning her the Bavarian Film Prize for Best Young Actress.
Jentsch's subsequent roles included the leading part in Hans Wilhelm Geißendörfer's "Schneeland" (2004). She continued to shine in Marc Rothemund's semi-documentary drama "Sophie Scholl - The Last Days" (2005),which premiered at the 55th Berlinale in February 2005. For her outstanding performance, Jentsch received the "Silver Bear" as best actress at the Berlinale. She also took home the award for Best Leading Role at the 55th German Film Awards in July 2005. "Sophie Scholl" went on to receive an Oscar nomination as a German contribution, solidifying Jentsch's reputation as a talented and versatile actress.