Jung Woo-sung debuted as an actor in the 1994 fantasy film The Fox with Nine Tails. In 1995, he starred in the SBS series Asphalt Man, which won him Best New Actor at the SBS Drama Awards and the 32nd Baeksang Arts Awards. In 1997, he reunited with his The Fox with Nine Tails co-star Go So-young in the action film Beat, in which he was named Best New Actor at the 17th Korean Association of Film Critics Awards. In 2003, he bagged a Best Actor nomination at the 24th Blue Dragon Film Awards for his performance in Mutt Boy. In 2008, he appeared in director Kim Jee-won’s The Good, the Bad, the Weird, which premiered at that year’s Cannes Film Festival. His work in the movie scored him the Best Supporting Actor citation at the 3rd Asian Film Awards. In 2011, he headlined the IRIS spin-off espionage series Athena: Goddess of War, which earned him a Top Excellence Award nomination at the SBS Drama Awards. In 2016 and 2018, he picked up Best Actor trophies for the movies Asura: The City of Madness and Steel Rain, respectively. In 2019, he and Kim Hyang-gi led the drama film Innocent Witness, which gave him numerous acting recognitions, including the Daesang at the 55th Baeksang Arts Awards and the 39th Golden Cinema Film Festival.

In 2011, Jung Woo-sung confirmed his relationship with his Athena: Goddess of War co-star Lee Jia. However, it was reported that the couple parted ways months after the controversy involving the actress’ secret marriage and divorce to South Korean music icon Seo Taiji broke out. In 2014, he became the first Korean to be appointed as goodwill ambassador of the United Nations High Commissioner for Refugees. Jung Woo-sung is close friends with his City of the Rising Sun co-actor Lee Jung-jae.