Robert De Niro Sr. was born in Syracuse, New York, to an Italian American father, Henry Martin De Niro (1897–1976), whose parents emigrated from Ferrazzano, in the province of Campobasso, Molise,[4] and an Irish American mother, Helen M. (née O'Reilly; 1899–1999). He was the eldest of three children; he and siblings John and Joan were raised in Syracuse, New York. De Niro studied at the renowned Black Mountain College under Josef Albers from 1939 to 1940. While Albers' highly analytical approach to painting did not appeal to De Niro's more instinctive style, the experience and international perspective of the Bauhaus master nonetheless left a lasting impression. De Niro studied with Hans Hofmann at his Provincetown, Massachusetts summer school.

After studying with Hans Hofmann in New York and Provincetown and Josef Albers at Black Mountain College, North Carolina in the late 1930s and early 1940s, De Niro worked for five years at Hilla Rebay’s legendary Museum of Non-Objective Art. From 1961 to 1964, De Niro traveled to France to paint in Paris and in the surrounding countryside.

Throughout the 1970s and 1980s, De Niro continued to exhibit in museums and galleries throughout the United States, including New York, San Francisco, Kansas City, Los Angeles, and Washington, D.C. He taught at several art schools and colleges including the Cooper Union, the New School for Social Research and the School of Visual Arts.

Robert De Niro Sr. died of cancer on the morning of May 3, 1993 (his 71st birthday) at his Manhattan home.