Jacobs was born in Ghent. As a child, he was a chorister at the Cathedral of Ghent. Later he studied classical philology at the University of Gent while continuing to sing in Brussels and in the the Hague.
The Kuijken brothers, Gustav Leonhardt and Alfred Deller encouraged him to pursue a career as a counter tenor, and he quickly became known as one of the best of his time. He recorded a large amount of less-known Baroque music of such composers as Cesti, d'India, Ferrari, Marenzio, Lambert, Guédron and others.
More recently, as a conductor, he has performed Orontea by Cesti, Xerse & Giasone by Cavalli, l'Incoronazione di Poppea and Il Ritorno d'Ulisse in Patria by Monteverdi, Flavio by Handel, Le Cinesi and Echo und Narziss by Gluck, La Finta Semplice by Mozart. Two recent, highly-praised recordings of Jacobs has been of Cosi Fan Tutte and Handel's Giulio Cesare
In 1992, the Berlin State Opera invited him to be the conductor of Cleopatra e Cesare by Graun, for the 250th anniversary of this work and of the Berlin opera.
As a conductor, René Jacobs regularly works with orchestras and ensembles such as the Concerto Köln, the Orchestra of the Age of Enlightenment, Akademie Für Alte Musik Berlin, Nederlands Kamerkoor and Rias-Kammerchor for recordings and concert tours, performing sacred music and oratorios.
In 1991, Jacobs was appointed artistic director of the opera programs at the Festwochen der Alten Musik in Innsbruck. He also teaches the interpretation and the Baroque singing style at the Schola Cantorum Basiliensis.