Hairston was born in Belews Creek, a rural community on the border of Stokes and Forsyth counties in North Carolina. At an early age he and his family moved to Pittsburgh, Pennsylvania. His grandparents had been slaves.
He graduated cum laude from Tufts University and studied music at the Juilliard School as well. He worked as a choir conductor in the early stages of his career. His work with choirs on Broadway eventually led to his singing and acting in plays, films, radio programs, and television shows.
In his later years, Hairston served as a cultural ambassador for American music, traveling to numerous countries with choral groups that he had assembled. In 1985 he took the Jester Hairston Chorale, a multi-racial group, to sing in China, at a time when foreign visitors were still quite rare in that country.
Hairston wrote the song "Mary's Boy Child" in 1956. He also wrote the song "Amen", which he dubbed for actor Sidney Poitier in the film Lilies of the Field (1963). He was a noted arranger of traditional "Negro spirituals" and choral director.
Most of Hairston's film work was in the field of composing, arranging, and choral conducting. Hairston also acted in over twenty films, mostly in small parts, some of which were not credited to him. Among the films he appeared in were "St. Louis Blues" (1958), The Alamo (1960), To Kill a Mockingbird (1962), and In the Heat of the Night (1967).
Hairston appeared on The Amos 'n' Andy Show. He had been in the radio program that was the basis for the TV show. He also played the role of Wildcat (1974-1975) on the show That's My Mama. In his senior years he appeared in the show Amen as Rolly Forbes (1986-1991). His last television appearance was in 1993 in the program Family Matters.
Hairston died January 18, 2000 of natural causes in Los Angeles, California, just six months shy of his 99th birthday.