Vladimir Ussachevsky was born in 1911 in the Russian province of Outer Manchuria, where his father was serving as a career officer in the Russian Army. Ussachevsky's mother performed and gave lessons on the piano, and she became her son's first teacher, initiating his musical education in piano and Russian Orthodox choral music. After playing in restaurants and vaudeville theaters as a teenager in Manchuria, Vladimir emigrated to California with his mother and siblings in 1930. Although he had intended to study electrical engineering, he became fascinated with composition while at Pomona College, where he received his B.A. in 1935. Ussachevsky then pursued graduate studies at the Eastman School of Music in Rochester, New York, studying with Howard Hanson among others. Ussachevsky returned briefly to California in the early 1940s after receiving his Ph.D. to care for his ailing mother. He taught high-school and junior-college music classes there before enlisting in the United States Army in September 1942 for the duration World War II.

After the war Ussachevsky began his long association with Columbia University, coming to New York in the fall of 1947 as an instructor and post-doctoral student under Otto Luening. At first, he taught introductory courses and continued to compose chamber, piano, and choral works in what he later called a "pseudo-Romantic Russian style." Ussachevsky's longstanding interest in electrical engineering, however, led him to pursue a radical new direction in composition. As his student Robert Moog recalled"The department had acquired its first tape recorder, an Ampex 400, and Vladimir was assigned to care for it. After using it extensively to record live performances, he began looking for new ways to use the tape machine. He created new musical sounds by speed changing, playing segments of tape backwards, splicing, looping, and electronic processing, and then assembled the sounds into experimental compositions." His tape compositions, many of them collaborations with Luening, were first performed in 1952, garnering Ussachevsky acclaim among both the New York musical community and the emerging transnational network of electronic music composers.

Inspired by his contacts with Pierre Schaefer, the creator of musique concrete, and electronic music composers in Russia, Ussachevsky along with fellow composers Luening, Milton Babbitt, and Roger Sessions founded in 1959 the Columbia-Princeton Electronic Music Center (CPEMC), which was modeled on Schaefer's Centre d'Etudes Radiophoniques in Paris. According to Moog, himself a major innovator in the field, Ussachevsky and engineer Peter Mauzey built equipment and developed technical practices of the highest possible quality, assisted by the funding of the Rockefeller Foundation and technical support from Bell Laboratories. Until his retirement from Columbia in 1980, Ussachevsky directed the CPEMC and mentored hundreds of students in theory and composition, most notably Moog and the composer Wendy Carlos. Ussachevsky also became a tireless crusader in the promotion of electronic music, presenting lecture-demonstrations at colleges and universities across the United States and Canada.

Outside of Columbia, Ussachevsky continued actively to pursue a composing career, writing scores of works for orchestras and choral groups, almost all of them incorporating the tape medium. Ussachevsky contributed musical settings for film, television, theater and poetry, collaborating with Clifton Fadiman, John Houseman, Stan Brakhage, Louis Zukofsky, Orson Welles, Alfred Hitchcock, and Burgess Meredith among others. He also played an active role in several artist organizations, principally the American Composers' Alliance and the MacDowell Colony, and served as editor for the journal New Music Edition.

After retiring from Columbia in 1980, he remained as a tenured professor at the University of Utah, where he had been Composer-in-Residence since 1970. He was married to the poet Elizabeth ("Betty") Kray from 1947 until her death in 1987. The couple had no children. Ussachevsky died on January 4, 1990 in New York City, remembered as a seminal figure in the development of electronic music in the United States.