Reared in Boston, Berenson was educated at Harvard University, from which he was graduated in 1887. His first book, The Venetian Painters of the Renaissance (1894), displayed a concise writing style. He was also endowed with a discriminating eye, exceptional memory, perceptive intelligence, and humanistic learning. For a time he was an adviser to the international art dealer Lord Duveen, and his opinion was often sought in the purchase of paintings. Many masterpieces now in American museums were bought upon his recommendation.

Although Berenson retained his U.S. citizenship, he lived in Italy most of his life. He was sequestered during World War II in Tuscany, and his diary Rumour and Reflection, 1941–1944, was published in 1952. He bequeathed to Harvard University his villa, I Tatti, with its art collection and magnificent library to be administered as a Center for Italian Renaissance Culture. Among his major works are Aesthetics and History in the Visual Arts (1948), The Drawings of the Florentine Painters (1938), and the monumental Italian Painters of the Renaissance (1952).