Born in British India, Khorana served on the faculties of three universities in North America. He became a naturalized citizen of the United States in 1966, and received the National Medal of Science in 1987.
Har Gobind Khorana was born to Indian parents Shrimati Krishna Krishna Devi Khorana and Shri Ganpat Rai Khorana in Raipur, a village in Punjab, British India at the time, which later became Pakistan. The exact date of his birth is not certain but he believed that it might have been 9 January 1922; this date was later shown in some documents, and has been widely accepted. He was the youngest of five children. His father was a patwari, a village agricultural taxation clerk in the Indian government. In his autobiography, Khorana wrote this summary: "Although poor, my father was dedicated to educating his children and we were practically the only literate family in the village inhabited by about 100 people." The first four years of his education were provided under a tree, a spot that was, in effect, the only school in the village.
He attended D.A.V. High School in Multan, in West Punjab, an area that is now in Pakistan. Later, he studied at the Punjab University in Lahore, with the assistance of scholarships, where he obtained a bachelor's degree in 1943 and a Master of Science degree in 1945.
Khorana lived in British India until 1945, when he moved to England to study organic chemistry at the University of Liverpool on a Government of India Fellowship. He received his PhD in 1948 advised by Roger J. S. Beer. The following year, he pursued postdoctoral studies with Professor Vladimir Prelog at ETH Zurich in Switzerland. He worked for nearly a year on alkaloid chemistry in an unpaid position.
During a brief period in 1949, he was unable to find a job in his original home area in the Punjab, which was in Pakistan by that time. He returned to England on a fellowship to work with George Wallace Kenner and Alexander R. Todd on peptides and nucleotides. He stayed in Cambridge from 1950 until 1952.