CCNY Knockout Huh-7 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human hepatocellular carcinoma Huh-7 cell line, engineered for targeted disruption of the CCNY gene. This polyclonal pool provides a heterogeneous loss-of-function model for investigating Cyclin Y-dependent signaling pathways without the need for single-cell cloning. The product is suitable for applications requiring a pooled knockout background, enabling robust analysis of gene function in a liver cancer cell context.
Huh-7 cells were established from a liver tumor of a 57-year-old Japanese male and are widely employed as an epithelial model of hepatocellular carcinoma. They retain hepatocyte-lineage features and are commonly used in liver cancer biology, virology, and drug metabolism studies. Their tumorigenic properties and well-characterized signaling landscape make Huh-7 an appropriate host for interrogating oncogenic pathways, particularly those dysregulated in hepatic malignancies.
Cyclin Y (CCNY) is a regulatory cyclin that partners with CDK14 (PFTK1) to phosphorylate the Wnt co-receptor LRP5/6 downstream of Wnt ligands such as WNT3A. This phosphorylation event potentiates ??-catenin stabilization by disrupting the destruction complex (AXIN, GSK3??, APC), leading to nuclear accumulation of ??-catenin and transcriptional activation of TCF/LEF target genes including MYC and CCND1. Cyclin Y also influences actin cytoskeleton organization and cell cycle progression, integrating signals from growth factor and E2F transcriptional regulators. The Cyclin Y?CCDK14?CLRP5/6 axis thus amplifies Wnt/??-catenin signaling, promoting proliferation and migration in Wnt-responsive cancer cells.
In the context of hepatocellular carcinoma, aberrant Wnt/??-catenin activation is a frequent oncogenic driver, making CCNY a relevant target for functional studies in Huh-7 cells. Disruption of Cyclin Y in this liver cancer model permits dissection of its contribution to CDK14-dependent LRP5/6 phosphorylation, ??-catenin stabilization, and downstream transcriptional programs. By comparing the polyclonal knockout population with wild-type Huh-7 cells, researchers can assess how loss of Cyclin Y affects tumor cell proliferation, migration, and Wnt pathway responsiveness, providing insight into potential therapeutic vulnerabilities in Wnt-driven liver malignancies.
This polyclonal knockout product is suited for diverse experimental workflows, including Western blotting and RT-qPCR to validate target gene disruption and pathway alterations, TOPFlash luciferase reporter assays to measure Wnt transcriptional activity, MTT proliferation assays to evaluate growth effects, and Transwell migration assays to assess metastatic potential. Additional applications include co-immunoprecipitation to study Cyclin Y interactors, immunofluorescence for ??-catenin localization, and flow cytometry for cell cycle profiling. The model is also valuable for drug screening campaigns targeting Wnt-driven hepatocellular carcinoma. For further details or technical assistance, please contact Ascent Research.
