James Rosenquist was born November 29, 1933 in Grand Forks, North Dakota. He grew up as an only child. His parents moved from town to town to look for work, finally settling in Minneapolis. His mother, who was also a painter, encouraged her son to have an artistic interest. In junior high school, Rosenquist won a short-term scholarship to study at the Minneapolis School of Art and subsequently studied painting at the University of Minnesota from 1952 to 1954. In 1955, at the age of 21, he moved to New York City on scholarship to study at the Art Students League.

From 1957 to 1960, he earned his living as a billboard painter. This was perfect training, as it turned out, for an artist about to explode onto the pop art scene. Rosenquist deftly applied sign-painting techniques to the large-scale paintings he began creating in 1960. Like other pop artists, Rosenquist adapted the visual language of advertising and pop culture (often funny, vulgar, and outrageous) to the context of fine art. Rosenquist achieved international acclaim in 1965 with the room-scale painting F-111.
Rosenquist has stated the following about his involvement in the Pop Art movement: "They(art critics) called me a Pop artist because I used recognizable imagery. The critics like to group people together. I didn't meet Andy Warhol until 1964. I did not really know Andy or Roy Lichtenstein that well. We all emerged separately."

His specialty is taking fragmented, oddly disproportionate images and combining, overlapping, and putting them on canvases to create visual stories. This can leave viewers breathless, making them consider even the most familiar objects (a U-Haul trailer, or a box of Oxydol detergent, etc.) in more abstract and provocative ways.

In addition to painting, he has produced a vast array of prints, drawings and collages. One of his prints, Time Dust (1992), is thought to be the largest print in the world, measuring approximately 7 x 35 feet.
In 1994 he created the print Discover Graphics in celebration of a Smithsonian educational program detailing the printmaking process. The print's proceeds supported the Smithsonian Associates' cultural and educational programs, and an original of the lithograph hangs in the Smithsonian Art Collectors Program's ongoing exhibition, Graphic Eloquence in the S. Dillon Ripley Center in the National Mall.

On April 25, 2009, a fire swept through Hernando County, Florida, where Rosenquist has lived for 30 years, burning the artist's house, studios, and warehouse. All of his paintings stored on his property were destroyed, including art for an upcoming show.