Clear cell renal cell carcinoma (ccRCC), a prevalent malignant tumor, exhibits resistance to chemotherapy, necessitating advanced therapeutic approaches. Sonocatalytic therapy has emerged as a promising non-invasive treatment for solid tumors due to its ability to penetrate deep tissue. Herein, the development of single atomic Au-doped 2D TiO2 nanosheets are presented with oxygen vacancies (Au-SA/Def-TiO2, ASDT) for sonocatalytic treatment of ccRCC. Surface defects on TiO2 nanosheets effectively stabilize Au single atomic sites via the Ti-Au-Ti structure, enhancing catalytic properties and facilitating reactive oxygen species (ROS) generation by reducing energy barriers and improving electron (e(-)) and hole (h(+)) separation. Additionally, ASDT exhibits peroxidase-like activities, further augmenting ROS production, leading to significant inhibition of tumor growth in subcutaneous and orthotopic ccRCC models under ultrasound irradiation. This study underscores the efficacy of defect engineering for stabilizing single atoms to boost ROS production and achieve effective tumor eradication by the enhanced sonocatalytic effect.