The ATG3 Knockout CAL-27 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population in which the ATG3 gene has been disrupted in the CAL-27 host cell background. This loss-of-function model is generated using CRISPR/Cas9-mediated gene targeting, resulting in a heterogeneous population of ATG3-null cells derived from the parental CAL-27 cell line. The product is supplied as a polyclonal pool, suitable for studies in which complete ablation of ATG3 function is required across a diverse genetic background, enabling robust assessment of autophagy-related phenotypes in an oral squamous cell carcinoma context.
CAL-27 is a well-characterized epithelial cell line originally established from a human tongue squamous cell carcinoma. As a model of oral squamous cell carcinoma, CAL-27 cells exhibit hallmark features of malignant epithelial tumors, including invasive and migratory capacities, and are widely employed in cancer research. The CAL-27 cell line provides a clinically relevant platform for investigating the molecular mechanisms driving head and neck cancers, particularly those involving autophagy-dependent survival pathways.
ATG3 encodes an E2-like enzyme essential for the autophagy conjugation cascade. Functioning downstream of the ATG7 E1-like enzyme and the ATG12?CATG5 complex, ATG3 catalyzes the covalent attachment of phosphatidylethanolamine (PE) to the C-terminal glycine of the LC3 family proteins, such as MAP1LC3B and GABARAP, generating the lipidated LC3-II form. This lipidation event is critical for autophagosome membrane elongation and closure. ATG3 activity is tightly regulated by upstream nutrient-sensing pathways, including mTORC1 and AMPK, as well as the transcription factor TFEB. The protein interacts directly with ATG7, the ATG12?CATG5 conjugate, ATG16L1, and WIPI2 to execute its function. Loss of ATG3 impairs the conversion of LC3-I to LC3-II, thereby blocking autophagic flux and leading to accumulation of cargo receptors such as SQSTM1/p62.
In the CAL-27 oral squamous cell carcinoma model, disruption of ATG3-mediated autophagy has profound implications for tumor cell biology. Autophagy often serves as a pro-survival mechanism in cancer cells coping with nutrient deprivation, hypoxia, and therapeutic stress; accordingly, ATG3 knockout in this background can be leveraged to dissect autophagy-dependent chemoresistance, metabolic adaptation, and invasive behavior. The combined ATG3 deficiency and CAL-27 tumorigenicity make this polyclonal population a powerful tool for probing the role of autophagy in oral cancer progression and for testing anti-cancer strategies that target autophagic pathways.
This product is suited for a broad array of functional experiments. Researchers can employ western blotting to monitor LC3-II and p62 levels as readouts of autophagy impairment, perform autophagy flux assays using lysosomal inhibitors, and visualize LC3 puncta via immunofluorescence. Additional applications include cell viability and clonogenic survival assays under nutrient starvation, as well as migration and invasion assays to evaluate metastatic potential. These cells also support dissection of upstream regulatory cascades involving mTOR, AMPK, and TFEB signaling. For further technical information and ordering details, please contact Ascent Research.
