The ATG3 Knockout A2780 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the A2780 human ovarian carcinoma cell line, designed to disrupt the ATG3 gene. This knockout model provides a loss-of-function system for studying autophagy mechanisms, with the polyclonal format ensuring a heterogeneous mixture of edited cells, reducing clonal artifacts. The use of CRISPR/Cas9-mediated gene disruption eliminates ATG3 expression, abolishing its critical E2-like enzymatic activity in the autophagy conjugation pathway.
The A2780 parental cell line is a widely used model of human epithelial ovarian carcinoma of the endometrioid subtype, established from an untreated patient. These cells retain key oncogenic signaling features and are extensively employed in ovarian cancer research, including studies of drug resistance, proliferation, and metastasis. As an adherent line with a robust growth profile, A2780 cells offer a reproducible platform for investigating the interplay between autophagy and ovarian cancer pathogenesis.
ATG3 functions as an E2-like enzyme essential for autophagosome formation, acting downstream of ATG7 and the ATG12-ATG5-ATG16L1 complex. It specifically catalyzes the covalent conjugation of phosphatidylethanolamine (PE) to the C-terminal glycine of LC3 family proteins (MAP1LC3A, MAP1LC3B, MAP1LC3C) and GABARAP subfamily members (GABARAP, GABARAPL1, GABARAPL2). This lipidation event facilitates LC3 recruitment to expanding autophagosomal membranes, a prerequisite for cargo sequestration and subsequent fusion with lysosomes. ATG3 activity is regulated by upstream nutrient-sensing pathways, including mTORC1 suppression and AMPK activation under starvation or hypoxia, thereby integrating autophagy induction with cellular metabolic status.
In the ovarian cancer context, autophagy exhibits dual roles in tumor suppression and promotion, making ATG3-mediated lipidation a critical node for therapeutic targeting. The ATG3 knockout in A2780 cells enables dissection of autophagy-dependent survival mechanisms under stress conditions such as nutrient deprivation or chemotherapeutic challenge. Because ovarian cancer cells often exploit autophagy for drug resistance, this polyclonal knockout population permits investigation of ATG3-dependent vulnerabilities and identification of synthetic lethal interactions with DNA-damaging agents or targeted therapies commonly used in ovarian cancer treatment.
Researchers can employ this model in a range of autophagy-focused assays, including western blotting to assess LC3 lipidation status, immunofluorescence for LC3 puncta formation, and autophagic flux measurements using lysosomal inhibitors. The system is also suited for cell viability assays under starvation or hypoxia, drug sensitivity screening, and RT-qPCR profiling of autophagy-related genes. Additionally, it serves as a valuable resource for functional genomics studies aiming to characterize the autophagy signaling network and for evaluating the impact of ATG3 loss on mitophagy and protein degradation. For further details on product usage and ordering, please contact Ascent Research.
