The ATG16L1 Knockout SK-HEP-1 Polyclonal Cells comprise a heterogeneous CRISPR/Cas9-edited population of SK-HEP-1 cells with disrupted ATG16L1, serving as a loss-of-function model for autophagy studies. These polyclonal knockout cells provide a robust model without clonal artifacts, supplied as a cryopreserved vial ready for expansion.
Derived from a male patient with hepatocellular carcinoma, the SK-HEP-1 cell line exhibits epithelial morphology and hepatocyte-like features, making it a relevant hepatic model for liver cancer research. Its tumor origin also allows exploration of autophagy??s dual tumor-suppressive and pro-survival roles.
ATG16L1 is a core component of the ATG12-ATG5-ATG16L1 complex, essential for LC3-I lipidation to LC3-II and autophagosome elongation. This complex is regulated by mTORC1, AMPK, and the ULK1 kinase complex, with innate immune signals such as NOD2 also modulating its assembly. ATG16L1 interacts with WIPI2 and FIP200 at the isolation membrane, working with ATG12-ATG5 conjugates to attach LC3 and GABARAP proteins to the phagophore. Consequently, ATG16L1 disruption blocks LC3 lipidation and autophagosome formation, halting canonical autophagic flux.
In hepatic cells, ATG16L1-mediated autophagy is vital for organelle quality control, mitophagy, and lipid metabolism, processes often aberrant in hepatocellular carcinoma. Knockout in SK-HEP-1 cells abolishes canonical autophagy, leading to accumulation of ubiquitinated cargo and mitochondrial damage under stress, facilitating studies on autophagy??s role in liver cancer survival, drug resistance, and metabolism. Additionally, given ATG16L1??s association with Crohn??s disease, this model can yield tissue-specific insights into autophagy deficiency.
This knockout cell model supports diverse applications: Western blotting to verify ATG16L1 loss and monitor LC3-II turnover with bafilomycin A1, immunofluorescence for LC3 puncta and p62 accumulation, and co-immunoprecipitation of complex partners. Flow cytometry, viability assays under starvation, and RT-qPCR enable quantitative autophagy and expression analysis. It is also valuable for screening autophagy modulators targeting liver cancer and inflammatory diseases. For further details, please contact Ascent Research.
