James Cronin was born on September 29, 1931. His father, James Farley Cronin, was a graduate student of classical languages at the University of Chicago. After his father had obtained his doctorate the family first moved to Alabama, and later in 1939 to Dallas, Texas, where his father became a professor of Latin and Greek at Southern Methodist University. After high school Cronin stayed in Dallas and obtained an undergraduate degree at Southern Methodist University in physics and mathematics in 1951.

While in graduate school he also met his wife, Annette Martin, whom he married in 1954. She was the Director of Special Events at the University of Chicago. They have three children: two daughters, Cathryn (1955) and Emily (1959), and a son, Daniel (1971). In June 2005 Annette Martin died of complications of Parkinson's disease. She was 71.

After obtaining his doctorate in 1955, Cronin joined the group of Rodney Cool and Oreste Piccioni at Brookhaven National Laboratory, where the new Cosmotron particle accelerator had just been completed. There he started to study parity violation in the decay of hyperon particles. During that time he also met Val Fitch, who brought him to Princeton University in Fall 1958. After Cosmotron underwent magnet failure, Cronin and the Brookhaven group moved to Bevatron at the University of California, Berkeley during the first half of 1958. Cronin and Fitch studied the decays of neutral K mesons, in which they discovered CP violation in 1964. This discovery earned the duo the 1980 Nobel Prize in Physics.

After the discovery, Cronin spent a year in France at the Centre d'Études Nucléaires at Saclay. After returning to Princeton he continued studying the neutral CP violating decay modes of the long-lived neutral K meson. In 1971, he moved back to the University of Chicago to become a full professor. This was attractive for him because of a new 400 GeV particle accelerator being built at nearby Fermilab.

Cronin received the Ernest Orlando Lawrence Award in 1976 for major experimental contributions to particle physics including fundamental work on weak interactions culminating in the discovery of asymmetry under time Reversal. In 1999, he was awarded the National Medal of Science.

When he moved to Chicago, he began a long series of experiments on particle production at high transverse momentum. With Physicist Pierre Piroue and colleagues we learned about many things. These are summarized in Physical Review D, vol 19, page 764 (1977). Following these experiments Cronin took a sabbatical at CERN in 1982–83, where he performed an experiment to measure of the lifetime of the neutral pion (Physics Letters vol 158 B page 81, 1985). He then switched to the study of cosmic rays. The first was a series of measurements looking for point sources of cosmic rays. No sources were found. A summary of the measurements was published in Physical Review D vol 55 page 1714 (1997). In 1998 he joined the faculty at the University of Utah on a half-time basis to work on ultra-high-energy cosmic ray physics and to jumpstart the Pierre Auger Observatory project. His appointment was to last five years, but he left after a year to continue gathering international support for the Observatory with Alan Watson and Murat Boratav.

Cronin was born in Chicago, Illinois, and attended Southern Methodist University in Dallas, Texas. He and co-researcher Val Logsdon Fitch were awarded the 1980 Nobel Prize in Physics for a 1964 experiment that proved that certain subatomic reactions do not adhere to fundamental symmetry principles. Specifically, they proved, by examining the decay of kaons, that a reaction run in reverse does not merely retrace the path of the original reaction, which showed that the interactions of subatomic particles are not invariant under time Reversal. Thus the phenomenon of CP violation was discovered.

In May 2011 his daughter Cathryn Cranston died of leukemia at age 54.