The NCOA4 Knockout AGS Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from the AGS human gastric adenocarcinoma cell line, carrying a targeted disruption of the nuclear receptor coactivator 4 (NCOA4) gene. This loss-of-function model provides a tool for investigating NCOA4-dependent biological processes, particularly ferritinophagy and cellular iron homeostasis. The polyclonal knockout format consists of a heterogeneous pool of edited cells, enabling population-level studies of gene function without clonal artifacts. It is suitable for assessing the overall impact of NCOA4 ablation on gastric cancer cell biology, including autophagy-related pathways and metal ion regulatory networks.
AGS cells are epithelial-like gastric adenocarcinoma cells originally isolated from a stomach adenocarcinoma, and they serve as a widely employed in vitro model for gastric cancer research. These cells retain key features of gastric mucosal epithelia, including malignant potential and responsiveness to oncogenic stimuli, making them relevant for studying tumor biology. Their use in autophagy and iron metabolism research is well established, owing to the critical role of iron in cancer cell proliferation and survival. The AGS background thus offers a pathologically relevant host for exploring how NCOA4-mediated processes influence gastric adenocarcinoma phenotypes.
NCOA4 functions as a selective autophagy receptor that binds ferritin, the major intracellular iron storage protein, and directs it to autophagosomes for lysosomal degradation in a process termed ferritinophagy. Mechanistically, NCOA4 interacts with ferritin heavy chain (FTH1) and light chain (FTL) subunits, as well as with LC3/GABARAP family proteins that mediate autophagosome cargo recruitment. Its activity is regulated by low intracellular iron levels and androgen receptor signaling, and it forms complexes with the ubiquitin ligase HERC2. The NCOA4 pathway converges with core autophagy components such as ATG7, LC3B, GABARAP, and the adaptor SQSTM1/p62, ultimately coupling ferritin turnover to the liberation of labile iron. NCOA4 knockout disrupts this delivery mechanism, leading to ferritin accumulation and impaired iron mobilization.
In the AGS gastric adenocarcinoma context, loss of NCOA4 impairs ferritinophagy flux, perturbing intracellular iron dynamics that are vital for sustaining malignant growth. Given that iron is a critical cofactor for enzymes involved in DNA synthesis, oxidative phosphorylation, and redox homeostasis, this knockout model offers insight into how iron dysregulation contributes to gastric cancer pathophysiology. It further enables dissection of the link between ferritinophagy and sensitivity to ferroptosis, a form of iron-dependent oxidative cell death that is increasingly implicated in tumor suppression and drug response. Thus, NCOA4 knockout AGS cells present a valuable platform for studying iron-addiction mechanisms in gastric adenocarcinoma.
This polyclonal knockout cell product is well suited for a range of experimental applications, including western blotting to measure ferritin and LC3 turnover, immunofluorescence colocalization of ferritin with autophagosomes, intracellular iron quantification using FerroOrange probes, and ferritinophagy flux assays in the presence of bafilomycin A1. It can be employed in RT-qPCR profiling of iron-responsive genes, cell viability assays under iron chelator treatment, and ferroptosis susceptibility testing via lipid peroxidation and cell death measurements. These applications allow researchers to probe the role of NCOA4 in gastric cancer iron metabolism, evaluate therapeutic strategies targeting ferritinophagy, and investigate the interplay between autophagy and oncogenic signaling. For further information or to request a detailed datasheet, please contact Ascent Research.
