Natalie Maines, Emily Robison and Martie Maguire will appear as the headlining act for July 2013's Craven Country Jamboree.
They will be replacing Lady Antebellum, who bowed out after singer Hillary Scott found out that she is pregnant.
Craven Country Jamboree is being held in Saskatchewan from July 11 to July 14 and will also feature performances by Tim McGraw, Gloriana, American Idol's Scotty McCreery and Kenny Chesney.
The Dixie Chicks' appearance at the festival will be their first concert since a benefit gig for wildfire relief in Texas in October 2011.
Maines and her bandmates have been on a recording hiatus since the release of their 2006 album Taking the Long Way, which earned them five Grammys.
They have sold more than 30 million albums worldwide since 1989.
Watch the Dixie Chicks perform 'Not Ready to Make Nice' below: