The DLGAP4 Knockout Huh-7 Polyclonal Cells product consists of a CRISPR/Cas9-edited polyclonal population of Huh-7 cells with targeted disruption of the DLGAP4 gene. This heterogeneous pool was generated by CRISPR/Cas9-mediated gene disruption, resulting in a loss-of-function model that retains genetic diversity of non-clonal engineered cells. Polyclonal knockout cells are ideal for population-level phenotypic studies, avoiding single-cell clone isolation. Individual cells may carry distinct editing events, and the pool presents a bulk knockout phenotype suitable for biochemical and cell-based assays.
The Huh-7 host cell line is an established hepatocellular carcinoma model from a well-differentiated tumor of a 57-year-old Japanese male. Huh-7 cells are extensively used in liver cancer research, including hepatocarcinogenesis, drug metabolism, and viral hepatitis (especially HCV). Their epithelial and tumorigenic nature makes them a standard platform for investigating oncogenic pathways and therapeutic vulnerabilities. Coupling Huh-7 with DLGAP4 knockout creates a dedicated tool for probing this scaffold protein??s role in liver cancer biology.
DLGAP4 encodes a scaffold protein linking membrane-associated guanylate kinases (MAGUKs) to the cytoskeleton. It interacts with DLG1/SAP97 and ??-catenin, contributing to cell adhesion complex assembly and Wnt/??-catenin signaling. DLGAP4 stabilizes ??-catenin, promoting its nuclear translocation and TCF/LEF-mediated transcription of oncogenes c-Myc and Cyclin D1. Upstream, DLGAP4 is regulated by the ??-catenin/TCF complex, miR-9, and EGF signaling. Downstream, it facilitates c-Myc transcription and cell cycle progression. Key pathway members include Wnt3a, Frizzled-7, Dishevelled-2, TCF4, and APC, converging on ??-catenin.
In hepatocellular carcinoma, DLGAP4 is increasingly recognized for its role in proliferation and invasion. Disruption of DLGAP4 in Huh-7 cells abrogates its scaffold interaction with DLG1 and ??-catenin, leading to ??-catenin destabilization and dampened Wnt/??-catenin signaling. This reduces transcription of c-Myc and expression of Cyclin D1, impairing cell cycle progression, migration, and invasive capacity. The resulting polyclonal knockout population enables investigation of DLGAP4??s oncogenic functions and its crosstalk with pathways such as Hippo signaling, which is implicated in liver cancer cell adhesion and polarity.
The DLGAP4 Knockout Huh-7 Polyclonal Cells support diverse functional assays. Researchers can examine Wnt/??-catenin pathway activity via TOPFlash/FOPFlash reporters, quantify protein and gene expression changes by Western blotting and RT-qPCR, and measure proliferation using CCK-8 or colony formation. Migration and invasion are assessed with Transwell assays, while co-immunoprecipitation and immunofluorescence reveal DLGAP4 interactors and ??-catenin localization. Applications extend to drug resistance and c-Myc regulatory network studies. For additional product information or technical support, please contact Ascent Research.
