Wnt/β-catenin signaling is a cellular signaling pathway that plays a crucial role in various biological processes, including embryonic development, cell differentiation, and tissue homeostasis. And, its imbalance is closely linked to tumorigenesis. Here, we report USP43, a deubiquitinase, as a positive regulator of Wnt/β-catenin signaling. Our study revealed that USP43 directly binds to β-catenin and stabilizes β-catenin through K48-linked deubiquitination, leading to the hyperactivation of downstream Wnt signaling. Functionally, USP43 promotes the migration and growth of colorectal cancer (CRC)and contributes to the survival of murine CRC organoids. These findings provide comprehensive understanding of USP43's impact on the Wnt/β-catenin pathway, offering further insights into colonic epithelial-related diseases.