NKU is an NCAA Division I school competing in the Horizon League, which it joined on July 1, 2015, after leaving the Atlantic Sun Conference.

[6] Prior to completing its transition to Division I, NKU changed its membership from the Atlantic Sun Conference to the Horizon League.

In 2000, the NKU women's basketball team became NKU's first national championship team by winning the NCAA Division II Women's Basketball Championship in overtime 71–62 over North Dakota State, ending its season with a 32–2 record.

[10] Dawn Plitzuweit, an assistant at Michigan, was named the new NKU Women's Basketball coach on May 10, 2012.

[14] In 2010, the NKU men's soccer team won the NCAA Division II national championship by defeating Rollins 3–2 in a driving snowstorm in Louisville.

[15] The team was led by senior Steven Beattie, who was named Ron Lenz National Player of the Year in both 2008 and 2010.

[18] Students have also organized club teams in ice hockey, taekwondo, fencing, boxing, lacrosse, rugby, kickball, skeet & trap, ultimate frisbee, and Brazilian jiu-jitsu.