Mwine was born in Hanover, New Hampshire to Ugandan parents in 1967. His father was a Harvard Law School-educated attorney. His parents separated when Ntare was 7, with Ntare spending time with his father (who was then working in finance in the United States, including a period at the World Bank in Washington D.C.) and his mother (who went to Kenya to teach psychology at the University of Nairobi).
Mwine earned a master's degree in Fine Arts from New York University's Graduate Acting Program at the Tisch School of the Arts, graduating in 1992. He also studied at the University of Virginia, the Moscow Arts Theatre, and the Royal National Theatre in London. Basing himself in Los Angeles, Mwine's first professional job was the role of Paul in the 1992 U.S. National Tour of Six Degrees of Separation, for which he received an NAACP Image Award nomination for Best Actor.
Mwine has also expressed his views on sticky subjects including LGBTQ , HIV/AIDS explaining: "I’m an artist. [I] have to tell stories”.

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