The 2000–01 Cypriot First Division was the 62nd season of the Cypriot top-level football league. Omonia won their 18th title.
Format
Fourteen teams participated in the 2000–01 Cypriot First Division. All teams played against each other twice, once at their home and once away. The team with the most points at the end of the season crowned champions. The last three teams were relegated to the 2001–02 Cypriot Second Division.
The teams had to declare their interest to participate in the 2001 UEFA Intertoto Cup before the end of the championship. At the end of the championship, the higher placed team among the interested ones participated in the Intertoto Cup (if they had not secured their participation in any other UEFA competition).
Point system
Teams received three points for a win, one point for a draw and zero points for a loss.
Before the start of the season, APOP Paphos and Evagoras Paphos were merged forming AEP Paphos, which took the place of APOP Paphos in the Cypriot First Division.