The DIP2A Knockout Huh-7 Polyclonal Cells product comprises a pool of CRISPR/Cas9-edited Huh-7 hepatocellular carcinoma cells with targeted disruption of the DIP2A gene locus, generating a heterogeneous polyclonal population suitable for loss-of-function studies. This format preserves the phenotypic diversity of the host cell line while enabling robust assessment of gene function without the clonal artifacts associated with single-cell-derived knockout cell lines. The polyclonal knockout approach offers a practical model for studying DIP2A biology in a liver cancer context.
Huh-7 cells are a well-characterized adherent epithelial cell line derived from a human hepatocellular carcinoma. They are extensively used in hepatology research to study hepatocyte biology, liver metabolism, viral hepatitis (particularly hepatitis C virus infection), and hepatocellular carcinoma pathogenesis. The cells retain key hepatocyte features and are susceptible to HCV, making them a relevant model for investigating molecular drivers of liver cancer and host-pathogen interactions.
DIP2A (Disco-interacting protein 2 homolog A) encodes a transmembrane protein possessing DMAP1-binding and follistatin-binding domains, and it is proposed to act as a cell surface receptor for the glycoprotein follistatin. Upon follistatin binding, DIP2A modulates TGF-?? superfamily signaling, influencing both canonical SMAD-dependent and non-canonical pathways. Mechanistically, DIP2A interacts with follistatin and the TGF-?? receptor complex, and its engagement is regulated by upstream factors including TGF-??1 and miR-543. Downstream, DIP2A signaling promotes SMAD2/3 phosphorylation, activates the PI3K/Akt pathway, upregulates cyclin D1 expression, and stimulates the MAPK/ERK cascade, thereby driving cell proliferation and survival.
In the context of hepatocellular carcinoma, DIP2A is implicated in tumor growth and progression through its role in follistatin-mediated signaling. Disruption of DIP2A in Huh-7 cells ablates the receptor function for follistatin, leading to attenuated SMAD2/3 phosphorylation and reduced Akt activity, which together diminish pro-proliferative signals. Consequently, this knockout model enables investigation of how loss of DIP2A impairs hepatocellular carcinoma cell viability, migration, and response to TGF-?? pathway modulators, revealing therapeutic vulnerabilities.
This polyclonal knockout cell population is suitable for a range of functional assays. Researchers can use western blotting and phospho-specific antibodies to quantify changes in SMAD2/3 phosphorylation and Akt activation, RT-qPCR to assess expression of downstream targets such as cyclin D1, and cell proliferation or migration assays (e.g., MTS, wound healing, or Boyden chamber) to evaluate phenotypic consequences of DIP2A disruption. Co-immunoprecipitation experiments can examine follistatin-DIP2A interactions, while RNA-seq enables global transcriptomic profiling of DIP2A-dependent gene networks. The model is particularly valuable for drug screening campaigns targeting the follistatin-DIP2A axis in liver cancer and for uncovering context-dependent roles of DIP2A in TGF-?? and PI3K/Akt signaling. For additional product information or technical support, please contact Ascent Research.
