Dr Karlien du Plessis completed her medical degree in 1995 from University of Pretoria. She did her internship at Pretoria Academic Hospital. She has a postgraduate diploma as well as a masters in palliative medicine.

From 1996 - 2003 she worked as a locum at various practices in the Pretoria area. She was also involved in examining sexually abused children as part of a multi-disciplinary team at the RP clinic.

In 2002 she commenced practising at the oncology practice of Dr Alberts Partners Incorporated in Vereeniging. She is actively involved in patient care in the unit as part of the multi-disciplinary team that aims to holistically look after every patient.