The ATG3 Knockout Ca Ski Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population of the human cervical carcinoma cell line Ca Ski (Homo sapiens), engineered for targeted disruption of the ATG3 gene. This product delivers a mixture of edited cells, providing a versatile loss-of-function model for studying autophagy without clonal isolation. By ablating the E2-like conjugating enzyme ATG3, these cells allow researchers to dissect its essential role in autophagosome biogenesis within an oncogenic background.
The parental Ca Ski line, derived from a cervical carcinoma metastasis, is HPV-16-positive and displays adherent epithelial morphology. It is a standard model for human papillomavirus-driven oncogenesis and cervical carcinoma research. These cells retain integrated viral oncogenes and respond to autophagy-modulating stimuli, making them well-suited for investigating the intersection of viral pathogenesis and cellular degradation pathways.
ATG3 functions as the catalytic E2-like enzyme that conjugates phosphatidylethanolamine (PE) to the C-terminal glycine of ATG8 family members, including LC3A, LC3B, GABARAP, and GABARAPL1, after their priming by the cysteine protease ATG4B. This lipidation reaction depends on the E1 enzyme ATG7 and the E3-like complex formed by ATG12?CATG5?CATG16L1. Upstream, ATG3 activity is positively regulated by nutrient deprivation, mTORC1 inhibition, and AMPK-mediated ULK1 kinase activation. ATG3 knockout eliminates lipidated LC3?CPE and GABARAP?CPE, halting phagophore elongation and autophagosome closure, thereby disrupting autophagic cargo degradation and flux.
In cervical cancer, autophagy exhibits context-dependent roles, functioning in tumor suppression while also being co-opted by cancer cells to survive metabolic stress, hypoxia, and chemotherapy. The HPV-16-positive Ca Ski line provides a clinically relevant platform to probe how ATG3-dependent autophagy influences cisplatin resistance, anoikis evasion, and invasive potential. This polyclonal knockout system enables the exploration of autophagy??s contribution to HPV oncoprotein-driven malignancy, offering insights into therapeutic vulnerabilities.
Researchers can employ these cells to investigate autophagosome formation mechanisms, autophagy-mediated drug resistance, and high-throughput screening of autophagy modulators. Typical downstream assays include immunoblotting for LC3-II and p62, autophagic flux measurements with lysosomal inhibitors (e.g., chloroquine), immunofluorescence visualization of LC3 puncta, transmission electron microscopy for autophagic structures, and functional tests such as MTT viability, Annexin V apoptosis, and transwell migration/invasion assays. This targeted knockout resource enables precise examination of ATG3??s role in cervical cancer biology. For technical support or ordering information, please contact Ascent Research.
