The IFT74 Knockout SK-HEP-1 Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal knockout population targeting the IFT74 gene in the SK-HEP-1 human hepatocellular carcinoma cell line. This product provides a heterogeneous pool of edited alleles, facilitating robust loss-of-function studies without clonal artifacts.
SK-HEP-1 cells derive from a liver adenocarcinoma and display epithelial morphology; they are a widely employed model for hepatocellular carcinoma (HCC) despite a debated endothelial origin. These cells exhibit hallmark malignant characteristics, including tumorigenicity in vivo, making them a relevant platform for investigating ciliary signaling in liver cancer.
IFT74 is a core subunit of intraflagellar transport complex B (IFT-B), partnering with IFT81, IFT88, and related proteins to assemble primary cilia via kinesin-2 and dynein-2 motors. Disruption of IFT74 abolishes ciliogenesis, leading to impaired Hedgehog (Hh) signaling; SMO fails to accumulate properly, preventing activation of GLI transcription factors and expression of target genes such as PTCH1 and CCND1. Concurrently, Wnt/??-catenin signaling is perturbed, with altered ??-catenin stabilization reducing MYC and CCND1 transcription. IFT74 expression is regulated by ciliogenic transcription factors RFX2, RFX3, and FOXJ1, integrating upstream Notch inputs. Through these interactions, IFT74 couples extracellular cues to downstream transcriptional programs essential for cell proliferation and differentiation.
In SK-HEP-1 HCC cells, IFT74 knockout yields a cilia-deficient model enabling dissection of primary cilium functions in liver cancer. Since Hh and Wnt pathways are frequently dysregulated in HCC, this system can illuminate how cilia-dependent signaling influences tumor cell growth, migration, and drug response. Additionally, it provides a cancer-relevant context for studying ciliopathy mechanisms, including those underlying short-rib polydactyly syndromes and Jeune syndrome, where IFT74 mutations cause developmental defects.
Applications include Western blotting and RT-qPCR for confirming IFT74 knockout and pathway perturbations, immunofluorescence for ciliary markers (acetylated ??-tubulin, ARL13B), Gli-luciferase reporter assays, cell proliferation and migration assays, and transcriptome analysis by RNA-seq. The polyclonal format is well-suited for drug screening and functional genomics studies targeting ciliary signaling in HCC. For further information or custom applications, please contact Ascent Research.
