The ATG4C Knockout SK-HEP-1 Polyclonal Cells offer a CRISPR/Cas9-edited polyclonal knockout population with targeted disruption of the ATG4C gene, encoding an autophagy-regulating cysteine protease. Derived from the SK-HEP-1 parental line, this heterogeneous pool enables robust loss-of-function studies while avoiding clonal selection artifacts. It serves as a versatile tool for dissecting autophagy pathways in a liver cancer context.
SK-HEP-1 is a human hepatic adenocarcinoma cell line originally established from ascites of a patient with liver adenocarcinoma. Displaying epithelial morphology, it is widely adopted in hepatocellular carcinoma research to explore tumor biology, metastatic behavior, and pharmacological responses, providing a physiologically relevant hepatic background for functional genomic manipulation.
ATG4C is an ATG4-family cysteine protease that processes Atg8-family proteins (MAP1LC3A/B, GABARAP, GABARAPL1) by cleaving their C-termini for conjugation to autophagosomal membranes and later deconjugates them, recycling LC3 and controlling autophagosome maturation. Regulated by nutrient deprivation, mTORC1 inhibition, AMPK, and TFEB, ATG4C modulates autophagic flux and LC3-II levels. It interacts with LC3 and GABARAP proteins and functions within a pathway including ULK1 complex, PI3K class III, ATG12-ATG5-ATG16L, ATG7, ATG3, and ATG4 proteases, ultimately regulating lysosomal degradation.
Disruption of ATG4C in these polyclonal cells impairs autophagic processing and may attenuate stress-induced autophagic degradation, relevant to hepatocellular carcinoma progression. This model enables dissection of ATG4C-dependent contributions to LC3 conjugation/deconjugation cycles, autophagic flux, and tumorigenic phenotypes within a hepatic microenvironment.
Typical applications include functional studies of ATG4C in autophagy-related tumorigenesis, drug sensitivity screens, and pooled CRISPR screening. Compatible assays encompass Western blotting for LC3-II/p62, LC3 puncta immunofluorescence, autophagic flux assays with chloroquine, RT-qPCR, cell proliferation and apoptosis assays, Transwell assays, and RNA-seq. For additional product details, contact Ascent Research.
