John Randal Kleiser was born in Lebanon, Pennsylvania, the son of Harriet Kelly (née Means) and Dr. John Raymond Kleiser. He has two brothers. Kleiser attended Radnor High School.
As a freshman at the University of Southern California, he appeared in George Lucas' student film Freiheit. (Kleiser also lived in the house that Lucas was renting at the time.) Kleiser graduated in 1968. His award-winning Master's thesis film Peege launched his career and was selected for preservation by the United States Library of Congress National Film Registry in 2007.
Kleiser directed several television movies in the mid-1970s, including Dawn: Portrait of a Teenage Runaway (1975) and The Boy in the Plastic Bubble (1975), which starred John Travolta. Kleiser was tapped to direct his first feature film, the 1978 film Grease, in large part because of Travolta's recommendation based on their work together on The Boy in the Plastic Bubble.

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