Edinburgh actor Jamie Sives will take to the stage of the Royal Lyceum Theatre in the capital to portray the firebrand preacher.
Glory on Earth will explore the 16th century religious figurehead’s relationship with Mary, Queen of Scots.
The iconic Scottish monarch will be played by Rona Morison, who previously starred alongside Sives in The James Plays, the epic historical trilogy staged by the National Theatre of Scotland at the Edinburgh International Festival in 2014.
She is due to appear alongside Simon Pegg and Mark Rylance in Steven Spielberg’s new science fiction film, Ready Player One, next year.
The Royal Lyceum said Glory on Earth, which runs from 20 May to 10 June, would “chart the fatal dance between a charming young queen and an uncompromising old zealot as they battle for the hearts and souls of the people of Scotland.”
Playwright Linda McLean’s script is said to have been inspired by four real-life meetings between Mary and Knox, which are recorded in Knox’s History of the Reformation.
Born in the Lochend area of Edinburgh, Sives also worked in a paper mill and was a nightclub doorman in the city.
He decided to pursue a dream of becoming a professional acting by taking evening classes in drama and made his name with a starring role in the feature film drama Wilbur Wants To Kill Himself.
His TV appearances include Doctor Who, Waking the Dead, To The Ends Of The Earth and Outcasts, while film appearances include Hallam Foe, Valhalla Rising and In The Heart of The Sea.
He performed alongside Danish actress Sofie Grabol and Taggart star Blythe Duff in Rona Munro’s trilogy The James Plays, playing King James III.
Glory on Earth will be directed by playwright David Greig, who took over the running of the Royal Lyceum last year.
McLean said: “I can honestly say there isn’t a commission that’s made me happier in a very long time.
“Delving into the lives of John Knox and Mary, Queen of Scots leaves me at turns warmly compassionate and fiercely outraged. It’s an astonishing story.”