Greyfriars Kirk is home to the Church of Scotland congregation of Greyfriars Tolbooth and Highland. The kirk takes its name from the pre-Reformation Franciscan friary that stood nearby. It was the first church built in Edinburgh after the Reformation and opened in 1620.

The kirk has a significant place in Scottish history. In 1638 the National Covenant, a document of great importance in the history of Scotland, was presented and signed in front of the pulpit. In 1679, some 1,200 Covenanters were imprisoned in Greyfriars Kirkyard pending trial.

In the 19th century the minister, Dr Robert Lee, led a movement to reform worship, introducing the first post-Reformation stained glass windows and one of the first organs in a Presbyterian Church in Scotland.

The church is primarily a place of worship, prayer, music and the preaching of the Gospel. However, it is also used for occasional concerts, organ recitals and dramatic presentations.