Heifetz studied violin from age three and at six performed Felix Mendelssohn's Violin Concerto. At nine he entered the St. Petersburg (Russia) Conservatory, where he studied under the noted violinist and teacher Leopold Auer. His first Berlin appearance in 1912 led to an invitation from the celebrated conductor Arthur Nikisch to play Tchaikovsky's Violin Concerto with the Berlin Philharmonic. He toured Europe from age 12.

In 1917 he fled the Russian Revolution via Siberia. He made his American debut in 1917 at Carnegie Hall, New York City, and became a U.S. citizen in 1925. He toured widely in Europe, the Orient, the Middle East, and Australia. Heifetz transcribed works of Johann Sebastian Bach, Antonio Vivaldi, and the 20th-century French composer Francis Poulenc for the violin. He also commissioned violin concerti by modern composers, among them Sir William Walton and Louis Gruenberg. After 1962 he taught at the University of Southern California, Los Angeles, where the Heifetz Chair in Music was established in 1975.