A member of the House of Gonzaga, Giovanni Vincenzo Gonzaga was born in Palermo on December 8, 1540, the son of Ferrante Gonzaga and his wife Isabella di Capua.^[1] He was the younger brother of Cardinal Francesco Gonzaga and the nephew of Cardinal Ercole Gonzaga.^[1]

As a young man, his kinsman Guglielmo Gonzaga, Duke of Mantua called him to Mantua to assist him in the government of the Duchy of Mantua.^[1] There, he became a member of the Knights Hospitaller. He was also made Prior of Barletta.^[1]

Pope Gregory XIII made him a cardinal deacon in the consistory of February 21, 1578.^[1] He received the red hat and the deaconry of San Giorgio in Velabro on November 21, 1578.^[1] On December 19, 1583, he opted for the deaconry of Santa Maria in Cosmedin.^[1]

He participated in the papal conclave of 1585 that elected Pope Sixtus V.^[1] On December 18, 1585, he opted for the order of cardinal priests and his deaconry was raised pro illa vice to the status of a titular church.^[1] He opted for the titular church of Santi Bonifacio e Alessio on April 20, 1587.^[1] Pope Sixtus V named him auditor of causes for citizens of the Papal States.^[1]

He participated in the first papal conclave of 1590 that elected Pope Urban VII; in the second papal conclave of 1590 that elected Pope Gregory XIV; and in the papal conclave of 1591 that elected Pope Innocent IX.^[1]

He died in Rome on December 23, 1591.^[1] He was buried in his titular church of Santi Bonifacio de Alessio.^[1]