The 2016 North Queensland Cowboys season was the 22nd in the club's history and their first as defending premiers. Coached by Paul Green and co-captained by Johnathan Thurston and Matthew Scott, they competed in the NRL's 2016 Telstra Premiership. In the pre-season the Cowboys competed in the 2016 Auckland Nines tournament, reaching the quarter-finals. The team finished the regular season in 4th, losing in the preliminary final to the eventual premiers, the Cronulla Sharks.
The NRL Auckland Nines is a pre-season rugby league nines competition featuring all 16 NRL clubs. The Cowboys, whose side featured a returning Matthew Bowen, finished first in their pool before being eliminated by the Melbourne Storm in the quarter-finals.
The following players have played a representative match in 2016.