The ATG7 Knockout A-549 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the A-549 human lung carcinoma cell line, featuring targeted disruption of the ATG7 gene. Designed to abolish ATG7 protein function, this polyclonal model enables loss-of-function studies while preserving the genetic heterogeneity typical of tumor cell populations, offering a robust system for investigating autophagy-dependent mechanisms in a lung epithelial context.
A-549 cells are an established epithelial line from human lung carcinoma tissue, widely used as a model for alveolar Type II pneumocytes and lung adenocarcinoma. Their adherent morphology and relevance to non-small cell lung cancer make them particularly suitable for respiratory research, including studies on cancer biology, drug metabolism, and epithelial cell function.
ATG7 encodes an E1-like enzyme that is essential for autophagy, a degradation process for cytoplasmic components. ATG7 activates the ubiquitin-like proteins ATG12 and LC3/ATG8 family members, catalyzing their conjugation to ATG5 and phosphatidylethanolamine, respectively??critical steps for autophagosome elongation and maturation. It functions downstream of nutrient and stress sensors including mTORC1, AMPK, and PI3K/AKT, which regulate the ULK1 kinase complex. Key interacting partners include ATG12, ATG3, ATG10, LC3, WIPI2, and ATG16L1. By driving LC3-II lipidation and autophagosome formation, ATG7 links upstream signaling to autophagic execution.
In A-549 lung cancer cells, ATG7 knockout abrogates autophagic flux, leading to accumulation of damaged organelles and protein aggregates. This reveals context-dependent roles of autophagy in tumor cell survival, proliferation, and drug resistance. As lung adenocarcinoma cells often display altered autophagic activity, disrupting ATG7 helps dissect whether autophagy suppresses or promotes tumor growth under specific microenvironments, making this model valuable for testing autophagy-targeting therapies.
Researchers can utilize these cells for autophagy flux studies, employing LC3 lipidation western blotting, p62/SQSTM1 degradation assays, and flux analysis with lysosomal inhibitors such as bafilomycin A1 or chloroquine. Additional applications include immunofluorescence for LC3 puncta, electron microscopy of autophagic structures, cell viability assays under starvation or chemotherapeutic stress, co-immunoprecipitation of ATG7 complexes, and RT-qPCR analyses of autophagy-related genes. The model supports cancer drug screening, metabolic stress profiling, and mechanistic dissection of mTOR/AMPK signaling. For detailed technical specifications, protocols, and pricing, please contact Ascent Research.
