The IFT25 Knockout SK-HEP-1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal population of SK-HEP-1 cells with targeted disruption of the IFT25 gene. This heterogeneous knockout pool serves as a loss-of-function model for investigating IFT25-mediated cellular processes. By avoiding clonal isolation, the polyclonal format captures a range of editing events, enabling studies that better reflect the genetic diversity of tumor cell populations.
SK-HEP-1 is an adherent epithelial-like cell line derived from a human hepatic adenocarcinoma. It is a well-characterized liver cancer model that retains the capacity for primary cilium formation under defined conditions. This feature makes it particularly useful for examining the crosstalk between ciliary structure/function and hepatic tumor biology.
IFT25 is an essential subunit of intraflagellar transport complex B, which mediates anterograde trafficking within primary cilia. It is indispensable for cilium assembly and Hedgehog signal transduction. Upon pathway activation by SHH or IHH ligands, relieved inhibition of SMO leads to GLI transcription factor activation; IFT25 is required for this process. Loss of IFT25 disrupts the ciliary localization of signaling intermediates, thereby attenuating GLI1/2-driven expression of targets such as CCND1, BCL2, and PTCH1. IFT25 physically interacts with IFT20, IFT27, IFT81, and HSPB11, and its expression is controlled by RFX transcription factors.
In SK-HEP-1 cells, IFT25 knockout provides a tractable system to dissect the contribution of ciliary Hedgehog signaling to hepatocellular carcinoma pathology. Aberrant activation of this pathway promotes proliferation, survival, and metastatic behavior in liver cancer cells. The polyclonal knockout model can be exploited to assess how disruption of cilium-dependent signaling influences tumor cell dynamics, drug response heterogeneity, and potential compensatory mechanisms that arise in the absence of functional IFT25.
This polyclonal knockout cell population is suitable for ciliogenesis assays using acetylated tubulin immunofluorescence, Hedgehog reporter assays (e.g., GLI-luciferase), and functional readouts including proliferation and migration. It can be integrated with Western blotting and RT-qPCR for molecular validation. Applications range from mechanistic studies of ciliary signaling to drug sensitivity screens in liver cancer. For further details, please contact Ascent Research.
