The CCDC127 Knockout Huh-7 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Huh-7 hepatocellular carcinoma cell line. This pool carries a heterogeneous array of CCDC127 gene disruptions, providing a robust loss-of-function model for functional studies without clonal selection artifacts. It is suitable for immediate use in phenotyping assays and pathway analysis in a liver cell context.
Huh-7 is an adherent epithelial cell line isolated from a liver tumor of a 57-year-old Japanese male, widely employed to study hepatocyte biology, hepatocellular carcinoma (HCC), and drug metabolism. These well-differentiated cells express hepatocyte markers and retain hepatic functions, making them an appropriate model for examining ciliary signaling in liver pathology and oncogenesis.
CCDC127 is a centrosomal protein that localizes to centriolar satellites and promotes primary cilium assembly by interacting with SDCCAG8, PCM1, and CEP290. It facilitates intraflagellar transport (IFT) protein recruitment, enabling cilium-dependent Hedgehog signaling. In this pathway, ligand binding to PTCH1 derepresses SMO, leading to GLI transcription factor activation and expression of targets like GLI1 and PTCH1. CCDC127 loss disrupts ciliogenesis and attenuates Hedgehog transduction, potentially affecting ??-catenin crosstalk.
Primary cilia are present on cholangiocytes and some hepatocytes, and Hedgehog signaling regulates hepatic processes including fibrosis and regeneration. In HCC, aberrant Hedgehog activation is common. CCDC127 knockout in Huh-7 cells thus models ciliary dysfunction in a hepatocellular context, enabling investigation of how ciliogenesis defects influence tumor cell proliferation, migration, and signaling. This system allows dissection of centrosome?Ccilium?CHedgehog interactions in liver cancer.
The polyclonal knockout cells are amenable to immunofluorescence detection of ciliary markers (acetylated ??-tubulin, ARL13B), ciliogenesis assays, Western blotting for CCDC127, SDCCAG8, and GLI1, and RT-qPCR analysis of Hedgehog targets (GLI1, PTCH1). They support co-immunoprecipitation, cell cycle flow cytometry, migration/invasion studies, drug screening for Hedgehog inhibitors, and RNA-seq. For more information, contact Ascent Research.
