Josefine Preuß was born on 13th of January 1986 in Zehdenick in Brandenburg. From early on, she appeared in theatre plays in Potsdam and became known to a wider audience for her role as the conniving Anna Reichenbach in the children's TV series "Schloss Einstein" (2000-2003). Preuß studied at the acting school "Die Etage" in Berlin, but did not graduate. Her movie career started in 2003 with the role of Emilia in the youth drama "Jargo", in which she starred alongside Constantin von Jascheroff and Nora von Waldstätten. Since then, she has been cast for multiple movies and TV productions, among them Irma-Kinga Stelmach's "Afterhour" (2005) and Felicitas Darschin's fairy tale adaptation "Zwerg Nase" (2007). The role of cheeky Lena in the successful German TV series "Türkisch für Anfänger" (2005-2008) marked her break-through. In 2007, she was nominated for the German Comedy Award, a year later she received the Actor award of the Grimme-Preis on behalf of the ensemble. After two appearances in the popular German crime series "Tatort" in 2008 and 2009, she starred in a supporting role in Detlev Buck's "Rubbeldiekatz" (2011). That same year, she again played Lena in the movie version of the TV series "Türkisch für Anfänger", which was released in German cinemas in March 2012.

Preuß' portrayal of the lead character in Uli Edel's "Das Adlon. Eine Familiensaga" (2012), a three-part series about the famous Berlin hotelier family, earned her a nomination at the Bayerischer Fernsehpreis. She played the lead in TV period dramas such as "Die Pilgerin" (2013) and "Die Hebamme" (Part 1 and 2, 2014 and 2016) and she appeared again on the big screen in a supporting role as the ex-girlfriend of an overly jealous man in Anno Saul's comedy movie "Irre sind männlich" (2014). In the GDR comedy "Vorwärts immer!" (2017), Josefine Preuß played the main character's young daughter, who in 1989 wants to leave the GDR with a fake ID and sets in motion all sorts of daring and turbulent events.