Jo Jae-yoon made his screen debut in the 2003 comedy film Please Teach Me English. He has since appeared in countless TV series and movies, mostly playing supporting characters. In 2010, he became part of the hit action-thriller film The Man From Nowhere. In 2012, he was cast in the SBS series The Chaser, which earned him a Special Award nomination at the SBS Drama Awards. In 2013, he appeared in the award-winning comedy-drama film Miracle in Cell No. 7. In 2016, he acted alongside Song Joong-ki and Song Hye-gyo in the widely popular KBS2 series Descended From the Sun. His performance in the drama won him the Scene Stealer Award at the 2nd Scene Stealer Festival as well as an Excellence Award at the 9th Korea Drama Awards. In 2017, he joined the main cast of KBS2’s Mad Dog, which scored him a Best Supporting Actor nomination at the KBS Drama Awards. That same year, he starred in the movie The Outlaws. In 2021, he took home the New Stealer Award at the Hallyu Culture Awards for tvN’s Mouse.

Jo Jae-yoon studied theater at the Seoul Institute of the Arts. He married a shopping channel host in February 2015.