François d'Amboise was the son of Jean d'Amboise and the brother of Adrien d'Amboise (Bishop of Tréguier), and of Jacques d'Amboise (rector of the University of Paris). He studied rhetoric and philosophy at the College of Navarre (Paris); in 1568 he became regent to the second grade there and taught literature and philosophy.
In 1575 he was named Procureur de la nation. In 1581, he became the king's lawyer in the treasury, and in 1586, advocate general to the Grand Conseil. In 1589, Henri III granted him letters of nobility. In 1596, Henry IV named him maître des requêtes, and then, in 1604, Conseiller d'État. François d'Amboise became seigneur of Vezeul, Bourot, Neuillé-le Lierre, Brouard, Lespinière, La Huardière in Touraine, of Houvoy, Malnoue, Courserin, Plessis-Bourré, Hémery, and Baron of La Chartre-sur-le-Loir.
He was friends with Robert Garnier, Pierre Matthieu and Gilles Bourdin, and it was at the house the latter that he met other dramatists of the period, including Guillaume Le Breton, Odet de Turnèbe and Pierre de Larivey.
He died in Paris in 1619 and was buried in the Church of Saint-Paul-Saint-Louis.
François d'Amboise wrote a comedy in verse entitled the Néapolitaines (1584) and several works of poetry, including an elegy on the death of Anne de Montmorency, Élégie sur le trépas d'Anne de Montmorency (1568). He was also responsible for an edition of the works of Peter Abelard (1616).
- Regrets facétieux et plaisantes harengues funèbres sur la mort de divers animaux pour passer le temps et resveiller les esprits mélencoliques, non moins remplis d'éloquence que d'utilité et gaillardise, Paris, N. Bonfons, 1583.
- The Néapolitaines, comedy (1584)