The DIXDC1 Knockout Huh-7 Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell population with disrupted DIXDC1 gene expression. This heterogeneous loss-of-function pool serves as a versatile tool for investigating the scaffold protein??s role in Wnt signaling. By avoiding single-clone selection, the polyclonal model mitigates clonal artifacts and better represents the biological variability inherent in the Huh-7 hepatocellular carcinoma background.
Huh-7 is a well-differentiated hepatocellular carcinoma cell line established from a liver tumor of a Japanese male. It retains hepatocyte-like epithelial morphology and tumorigenic properties, and it harbors constitutively active Wnt/??-catenin signaling. This background offers a clinically relevant hepatic environment for studying DIXDC1-dependent phenotypes in liver cancer.
DIXDC1 encodes a scaffold that positively regulates canonical Wnt signaling by promoting Dishevelled (DVL2)-Axin (AXIN1) interaction, leading to ??-catenin stabilization. Upon WNT ligand stimulation, DIXDC1 bridges DVL2 and AXIN1, counteracting the destruction complex containing AXIN1, GSK3??, and APC. Stabilized ??-catenin translocates to the nucleus, associating with TCF/LEF factors to activate transcription of targets such as MYC, CCND1, AXIN2, LEF1, and TCF7. CRISPR/Cas9 disruption of DIXDC1 impairs this scaffolding function, attenuating ??-catenin accumulation and downstream gene expression.
In hepatocellular carcinoma, aberrant Wnt activation drives tumor progression, and DIXDC1 overexpression amplifies oncogenic signals. The Huh-7 knockout population enables dissection of DIXDC1??s contribution to HCC proliferation, migration, and invasion. As Huh-7 cells rely on Wnt signaling, this model provides a physiologically relevant system to study how loss of DIXDC1 rewires the network and alters cancer cell behavior.
Researchers can apply this knockout pool in assays such as western blotting for ??-catenin and phospho-??-catenin, TOPFlash/FOPFlash dual-luciferase reporter assays to measure TCF/LEF transcriptional activity, and RT-qPCR for Wnt target genes. Co-immunoprecipitation verifies disrupted DIXDC1-AXIN1 binding. Functional readouts include CCK-8 viability, wound healing migration, and Transwell invasion assays. The polyclonal nature suits high-throughput screening of Wnt pathway inhibitors and drug target validation in liver cancer biology. For further technical details, contact Ascent Research.
