The DVL3 Knockout Huh-7 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human Huh-7 hepatocellular carcinoma cell line, with targeted disruption of the DVL3 gene. This loss-of-function model enables study of DVL3 in Wnt signal transduction. The polyclonal pool preserves editing heterogeneity, avoiding clonal selection artifacts and providing a robust system for investigating DVL3-dependent processes. Researchers can interrogate both canonical and non-canonical Wnt pathways in a liver cancer context.
Huh-7 is a widely used adherent epithelial cell line from a hepatocellular carcinoma of a 57-year-old Japanese male. It retains hepatocyte features and serves as a model for liver cancer, enabling studies on proliferation, migration, and signaling. This background is particularly valuable for examining Wnt pathway aberrations common in hepatocellular carcinoma, where ??-catenin dysregulation drives tumorigenesis.
DVL3 is a cytoplasmic phosphoprotein that mediates signaling downstream of Wnt-activated Frizzled receptors and LRP5/6 coreceptors. Upon Wnt stimulation, DVL3 is phosphorylated by CK1 and PAR-1, recruiting the Axin/APC/GSK3?? complex to the membrane and stabilizing ??-catenin (CTNNB1). Nuclear ??-catenin partners with TCF/LEF factors like TCF7L2 to induce targets such as c-Myc and Cyclin D1. DVL3 also governs non-canonical Wnt/PCP signaling via RhoA and Rac1, and Wnt/Ca2+ pathways through JNK, regulating polarity, migration, and calcium flux. Key interacting partners include Frizzled receptors, Axin, APC, GSK3??, CK1, PAR-1, and ??-catenin.
In Huh-7 cells, DVL3 disruption impairs Wnt signal transmission, attenuating ??-catenin-driven transcription and non-canonical activities. This likely alters proliferation and migration balance in liver cancer. These polyclonal knockout cells thus provide a physiologically relevant model to dissect DVL3 contributions to hepatocarcinogenesis, separate canonical from non-canonical Wnt functions, and assess DVL3 dependency. The model also enables exploration of cross-talk with other oncogenic pathways active in Huh-7.
Applications include luciferase reporter assays (TOP/FOP flash), RT-qPCR for Wnt targets (c-Myc, Cyclin D1), Western blotting for DVL3 and ??-catenin, and immunofluorescence for ??-catenin localization. Functional assays cover proliferation (MTT, BrdU), migration, invasion, and co-immunoprecipitation of ??-catenin complex components. The cells are suitable for drug screening and canonical versus non-canonical pathway crosstalk studies. For further information, please contact Ascent Research.
