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In 1559, John Knox delivered his inaugural sermon on the Reformation at the High Kirk of St. Giles. His captivated audience noted that "he was so fervent and vigorous it seemed as if he might shatter the pulpit and soar from it." Knox played a pivotal role in disseminating the Presbyterian form of Protestantism across Scotland.

Fast forward to 1633, when King Charles I appointed Scottish Episcopal bishops. By 1635, William Forbes was consecrated as the first bishop of the newly established diocese of Edinburgh, transforming St. Giles' into a cathedral and the official seat of a bishop.

It was within the walls of St. Giles on 23 July 1637 that Jenny Geddes made history. As Rev. James Hannay began to read from Laud's Liturgy, she rose from her seat, seized a stool, and hurled it at him, exclaiming, “Villain, dost thou say mass at my lug (in my hearing)”. This act ignited a riot, with others joining in, shouting and throwing stools, before storming out of the building. This incident marked the beginning of the period of Covenanter oppression.

Although it is today a Presbyterian church, which does not have bishops, St. Giles' continues to be referred to as a cathedral.