Guise was the son of Claude of Guise and his wife Antoinette of Bourbon. His sister Mary of Guise was wife of James V of Scotland and mother of Mary I of Scotland.
He was made Archbishop of Reims in 1541 and Cardinal of Lorraine in July 1547.
Along with his brother Francis, Duke of Guise he was powerful in the government of France during the reign of Francis II from 1559 to 1560.
Guise was one of the leaders of the Catholic faction in the French Wars of Religion.