Born in Lucerne, Regula Mühlemann studied singing with Barbara Locher at the Hochschule Luzern - Muzik. She made her first appearances on the opera stage at the Luzerner Theater, which led to a performance at the Teatro La Fenice in Venice (Despina in Cosí fan tutte). In 2012 she made her mark at the Salzburg Festival in von Winter's opera Das Labyrinth. Regula Mühlemann has received numerous awards, and was a finalist in the 2015 Cardiff Singer of the World Competition. She has performed on the most prestigious stages such as Salzburg, Berlin, Paris, Zurich, Lucerne and Geneva.

Regula Mühlemann has performed the roles of Maturina in Gazzaniga's Don Giovanni Tenorio, Barbarina in The Marriage of Figaro, Papagena in The Magic Flute, and Doralice in Scarlatti's Il Trionfo dell'onore. She begins the 22/23 season as Euridice in Gluck's Orfeo ed Euridice under the musical direction of Thomas Hengelbrock at the Théâtre des Champs-Elysées in Paris and returns to Theater Basel with her role debut as Gilda in Verdi'sRigoletto.

Son deuxième album Cleopatra, enregistré en 2017, a également été salué par la critique : « how captivating beautiful does the voice alloyed with gold and silver » (Sound). En 2018, elle reçoit l’OPUS Klassik « Newcomer of the Year ».