The ATG7 Knockout SK-HEP-1 Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout cell population in which the ATG7 gene has been disrupted across a bulk population of SK-HEP-1 human hepatic adenocarcinoma cells. This pooled format provides a heterogeneous loss-of-function model without clonal selection, enabling the study of autophagy defects in a genetically diverse cellular context. The polyclonal nature reflects the aggregate effect of multiple editing events, making it suitable for applications that require population-level responses rather than clonal homogeneity.
SK-HEP-1 is an ascites-derived human hepatic adenocarcinoma cell line that serves as a well-established model for hepatocellular carcinoma research, liver cancer biology, and metabolic studies. Its hepatic origin and epithelial morphology allow investigation of autophagy-related processes in the context of liver disease, including lipid accumulation, fibrogenesis, and drug metabolism. The cell line retains key signaling pathways relevant to hepatic function, making it a valuable host for genetic manipulation aimed at dissecting autophagy-dependent phenotypes in liver malignancy and metabolic disorders.
ATG7 encodes an E1-like activating enzyme that is indispensable for autophagy, operating in two ubiquitin-like conjugation cascades. It activates ATG12 for transfer to ATG10, ultimately forming the ATG12?CATG5 conjugate, and activates LC3 family proteins (LC3A/B/C) and GABARAP for conjugation to phosphatidylethanolamine, a critical step for autophagosome membrane elongation. ATG7 activity is tightly regulated by upstream nutrient-sensing kinases, including mTORC1 and AMPK, and transcription factors such as TFEB and FOXO3. Downstream, ATG7-mediated lipidation of LC3B facilitates recruitment of cargo receptors p62/SQSTM1 and NBR1, promoting selective autophagy of ubiquitinated substrates and damaged mitochondria. Key interacting partners include ATG3, ATG5, ATG12, ATG16L1, and the LC3 conjugation machinery, positioning ATG7 at the core of autophagosome biogenesis.
In the SK-HEP-1 hepatic adenocarcinoma background, disruption of ATG7 abrogates autophagy-dependent cytoprotective and metabolic functions, yielding a model with direct relevance to liver cancer pathophysiology. Loss of autophagic flux leads to accumulation of dysfunctional mitochondria and protein aggregates, recapitulating aspects of hepatic steatosis, fibrosis, and ER stress responses. This system enables dissection of autophagy??s role in chemoresistance, metabolic reprogramming, and crosstalk with the PI3K/AKT/mTOR signaling axis, providing a platform to explore therapeutic vulnerabilities in ATG7-deficient hepatocellular carcinoma.
This polyclonal knockout pool is ideal for autophagy research applications such as autophagic flux assays with bafilomycin A1, Western blotting for LC3-II and p62, fluorescence microscopy for LC3 puncta, and co-immunoprecipitation of the ATG12?CATG5 conjugate. It is well-suited for high-throughput screening of autophagy modulators, cell viability studies under nutrient deprivation, and siRNA rescue experiments to confirm ATG7-dependent effects. For more information or custom services, please contact Ascent Research.
