The DVL2 Knockout Huh-7 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population optimized for functional analysis of the DVL2 gene in a liver cancer model. This product consists of a pool of Huh-7 human hepatocellular carcinoma cells harboring targeted disruptions of the DVL2 locus, introduced via CRISPR/Cas9-mediated gene editing. The polyclonal format yields a mixed cellular population with collective ablation of DVL2 function, providing a versatile loss-of-function system for pathway dissection without the need for clonal isolation.
The Huh-7 cell line was originally derived from a hepatocellular carcinoma of a 57-year-old Japanese male and is widely utilized as a model for liver cancer. Huh-7 cells maintain hepatocyte-like features and often exhibit constitutive activation of Wnt/??-catenin signaling, a hallmark of hepatocellular carcinoma. This background makes DVL2 knockout in Huh-7 particularly relevant for probing Wnt pathway dependencies in HCC.
DVL2 encodes a cytoplasmic scaffold that serves as a central mediator of both canonical Wnt/??-catenin and non-canonical planar cell polarity (PCP) signaling. Upon Wnt ligand stimulation of Frizzled receptors, DVL2 undergoes phosphorylation by CK1 and GSK-3?? and recruits the ??-catenin destruction complex??comprising Axin, APC, GSK-3??, and CK1??to the plasma membrane. This event results in stabilization and nuclear accumulation of ??-catenin, which partners with TCF/LEF transcription factors to activate Wnt target genes. In the PCP branch, DVL2 signals through Rho GTPases and JNK to regulate cytoskeletal dynamics and cell migration. DVL2 directly interacts with Axin, ??-catenin, CK1, GSK-3??, and DAAM1 to orchestrate these cascades.
In hepatocellular carcinoma, aberrant Wnt/??-catenin signaling frequently drives tumorigenesis, proliferation, and metastasis. Disruption of DVL2 in Huh-7 cells provides a controlled experimental system to dissect the contribution of Wnt signal transduction to liver cancer cell behavior. This knockout model enables researchers to distinguish between DVL2-dependent ??-catenin signaling and ??-catenin-independent effects, and to assess oncogenic pathway addiction in a clinically relevant HCC model. Moreover, it permits investigation of DVL2 functions beyond ??-catenin, such as its role in PCP-mediated cell migration and invasion.
These polyclonal knockout cells are well-suited for a range of functional assays, including TOP/FOP flash reporter assays for Wnt/??-catenin activity, Western blotting for ??-catenin stabilization and DVL2 phosphorylation, RT-qPCR for downstream target gene expression, and co-immunoprecipitation of DVL2 with Axin or CK1. They also support migration and invasion assays under differential Wnt pathway stimulation and drug sensitivity profiling with Wnt inhibitors. For additional information or assistance with custom applications, please contact Ascent Research.
