The IFT27 Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population bearing targeted disruption of the IFT27 gene. This loss-of-function model provides a heterogeneous pool of SK-HEP-1 cells with IFT27 gene ablation, suitable for studying intraflagellar transport (IFT) and hedgehog signaling without the need for clonal isolation.
The SK-HEP-1 host cell line, derived from ascites of a 52-year-old Caucasian male with hepatic adenocarcinoma, serves as a well-established model for hepatocellular carcinoma and metastasis. It exhibits an endothelial-like phenotype and is frequently employed to investigate tumor signaling, migration, and drug response.
IFT27 encodes a small GTPase that functions as a core subunit of the intraflagellar transport complex B (IFT-B), essential for retrograde ciliary trafficking and hedgehog signal transduction. IFT27 directly interacts with IFT25, IFT81, and the BBSome complex subunits BBS1, BBS4, BBS5, and BBS8, and operates downstream of Sonic hedgehog (SHH) ligand and the transcription factors RFX and FOXJ1. Within the cilium, IFT-B components including IFT20, IFT25, IFT81, and motors KIF3A and DYNCH1 facilitate the movement of signaling effectors such as SMO and GLI transcription factors. IFT27-dependent transport is required for GLI activator processing and transcriptional induction of hedgehog targets like PTCH1, HHIP, CCND1, and MYC. Loss of IFT27 disrupts ciliogenesis, alters SMO ciliary localization, and deregulates hedgehog target gene expression, thereby linking defective IFT to ciliopathy phenotypes and aberrant pathway activation in cancer.
In the context of SK-HEP-1 hepatic adenocarcinoma, IFT27 knockout enables dissection of the role of ciliary hedgehog signaling in liver cancer, where pathway dysregulation is commonly associated with tumor progression and drug resistance. This model recapitulates molecular features of ciliopathies such as Bardet-Biedl syndrome and skeletal ciliopathies, offering a unique platform for exploring the intersection of intraflagellar transport and hepatocellular carcinoma. The polyclonal knockout population reflects heterogeneous gene disruption, providing a more physiologically relevant system for signaling studies.
These cells can be utilized in Western blotting for IFT27, GLI1, and PTCH1; immunofluorescence for acetylated tubulin to visualize primary cilia; RT-qPCR for hedgehog target genes; RNA-seq for transcriptomic profiling; flow cytometry for cell cycle analysis; wound-healing assays for migration; and drug sensitivity testing with smoothened antagonists. Co-immunoprecipitation can assess IFT-B complex assembly, and PDGFR?? phosphorylation assays can monitor hedgehog pathway activity. Applications include ciliopathy modeling, hedgehog signaling investigation, ciliary transport analysis, and liver cancer drug screening. For further details, contact Ascent Research.
