Lee Joo-woo landed her first screen role in JTBC’s teen crime series Seonam Girls’ High Investigation Unit in 2014. The following year, she became part of KBS1’s two-part television special Snowy Road, which was later theatrically released as a drama film. In 2017, she appeared in the MBC daily series Return of Fortunate Bok, which earned her a Best Character Award nomination at the MBC Drama Awards. In 2018, she joined the main casts of JTBC’s Woohoo Waikiki and tvN’s Let’s Eat 3. In 2019, she acted alongside Lee Yo-won and Choi Gwi-hwa in the OCN series Running Investigators. In 2022, she was cast in the SBS legal drama Why Her.

Lee Joo-woo attended Baekseok Arts University’s music department.