ATG4C Knockout A-549 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the A-549 human lung adenocarcinoma cell line, designed to disrupt the ATG4C gene. This polyclonal pool provides a genetically diverse loss-of-function model that reflects heterogeneous knockout events, enabling robust study of autophagy-related processes without clonal artifacts. The product is supplied as a ready-to-use polyclonal cell population, validated to lack ATG4C protein activity, and is suitable for high-throughput screening and mechanistic studies.
The parental A-549 cell line originates from a 58-year-old male with lung carcinoma and is widely used as a model of alveolar type II pneumocytes. A-549 cells exhibit an epithelial morphology and harbor a KRAS G12S mutation while retaining wild-type p53, making them a relevant system for investigating lung adenocarcinoma biology and oncogenic signaling. This established line enables studies of epithelial-derived cancer cell behavior, including proliferation, migration, and drug response.
ATG4C encodes a cysteine protease that plays a critical role in the autophagy pathway by processing pro-LC3 proteins. Mechanistically, ATG4C cleaves the C-terminal tail of pro-LC3B, GABARAP, and GABARAPL1, exposing a glycine residue necessary for conjugation to phosphatidylethanolamine. This priming event, facilitated by ATG7 and ATG3, allows LC3 lipidation and incorporation into expanding autophagosomal membranes. ATG4C activity is regulated upstream by nutrient deprivation signals and the mTORC1-TFEB axis. Its disruption impairs key downstream events, including LC3B conversion and autophagosome formation, and affects interaction partners such as LC3B, GABARAP, and ATG7. Consequently, knockout of ATG4C leads to defective autophagy flux and accumulation of cargo adaptors like p62/SQSTM1.
In the context of A-549 lung adenocarcinoma cells, ATG4C knockout provides a powerful tool to dissect autophagy-dependent processes in cancer. Aberrant autophagy is implicated in tumor progression, therapy resistance, and metabolic adaptation. By eliminating ATG4C function, researchers can investigate how loss of a key autophagic protease influences A-549 cell viability, sensitivity to chemotherapeutics, and invasive potential. This model is particularly valuable for elucidating the role of autophagy in KRAS-driven lung cancers and for validating ATG4C as a therapeutic target.
Typical applications include western blotting to assess LC3B lipidation and p62 accumulation, immunofluorescence microscopy for LC3 puncta quantification, and autophagy flux assays using chloroquine to measure autophagic turnover. The polyclonal knockout population is well-suited for functional genomics screens, drug sensitivity profiling of autophagy modulators, and migration/invasion scratch wound healing assays. It also supports RT-qPCR analysis of transcriptional responses downstream of ATG4C disruption. Researchers may employ these cells to screen for ATG4C-specific inhibitors or to study autophagy??s role in lung adenocarcinoma progression. For further information on customization, licensing, or technical support, please contact Ascent Research.
