The son of an Ontario judge, Norman Mackenzie was born in Sarnia, Ontario, in 1869, and moved west to Regina in 1891. He became a well-known lawyer, an active member of the Regina community, and an avid art collector. He began collecting art in 1900 and the collection grew to include works he gathered during the annual trips that he took to Europe until 1914, works from the Far and Near East, and works by local western Canadian artists, such as Henderson and Sheldon-Williams. At his death in 1936, he left his art collection and a bequest to Regina College with the hopes that an art gallery would be established. The first curator for the Mackenzie collection was Augustus Kenderdine and an investigation into the construction requirements for such a building was begun as early as 1937.

The Norman Mackenzie Art Gallery and Art School was completed in 1952 and officially opened on September 25th, 1953. Under Regina Five member Ron Bloore, who followed upon the first curator, Richard Simmons, the gallery achieved national prominence. Exhibitions of note during this time included the controversial Win Hedore exhibition for the fictitious artist of that name, and the May show, which led to the 1961 National Gallery exhibition Five Painters from Regina.

In 1981, the position of coordinator of Education and Extension was created to administer community and outreach programmes previously undertaken by the Tour Committee of the independently run Norman Mackenzie Art Gallery Society. On May 1st, 1990, the gallery became independent of the University of Regina, which continued to hold two positions on the Board of Trustees. The gallery now operates as the MacKenzie Art Gallery Incorporated and has been located at 3475 Albert Street since August 1990.