The ATG3 Knockout AGS Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the AGS human gastric adenocarcinoma cell line, carrying targeted disruption of the ATG3 gene. This product provides a heterogeneous pool of gene-edited cells that circumvents clonal selection bias, offering a robust loss-of-function model for studying ATG3-dependent autophagy mechanisms. The polyclonal format enables experiments under conditions that better represent genetic variability, while the CRISPR/Cas9-mediated gene disruption ensures efficient ablation of ATG3 protein function without resort to monoclonal isolation. Researchers can employ this population to interrogate the roles of ATG3 in autophagosome biogenesis, cargo degradation, and cellular stress responses.
The AGS host cell line originates from a poorly differentiated human gastric adenocarcinoma, displaying adherent epithelial morphology. Widely adopted in gastric cancer research and Helicobacter pylori infection studies, AGS cells serve as a model for gastric epithelial biology, including proliferation, survival, and pathogen-host interactions. Their molecular toolkit includes activated oncogenic pathways commonly mutated in gastric cancers, making them particularly relevant for dissecting autophagy contributions to tumor maintenance. The cell line??s robust growth characteristics and reproducible behavior in standard culture conditions facilitate high-quality assays and consistent experimental outcomes.
ATG3 functions as an E2-like conjugating enzyme within the ubiquitin-like ATG8/LC3 lipidation system, a core mechanism for autophagosome elongation. Upon nutrient deprivation or mTORC1 inhibition, activated ULK1 and the Beclin-1/VPS34 complex initiate phagophore nucleation, allowing ATG7 (E1-like) to activate LC3 family proteins. ATG3 receives the activated LC3 and, in concert with the ATG12-ATG5-ATG16L1 E3-like complex, catalyzes covalent conjugation of LC3 to phosphatidylethanolamine on the expanding membrane. This drives membrane curvature and selective cargo recruitment through adaptors such as p62/SQSTM1 and NBR1. Upstream regulators including AMPK and ATG7, and downstream targets like MAP1LC3A/B, GABARAP, and GABARAPL1, place ATG3 at a convergent hub linking energy sensing to autophagic flux.
In gastric adenocarcinoma, ATG3-mediated autophagy often supports tumor cell survival under metabolic stress, promotes chemoresistance, and facilitates epithelial-mesenchymal transition. Disrupting ATG3 impairs LC3 lipidation and autophagosome completion, sensitizing cells to nutrient exhaustion and cytotoxic drugs. During H. pylori infection, the bacterium manipulates autophagy to create a replicative niche; ATG3 knockout can alter bacterial persistence and host inflammatory responses, yielding insights into chronic gastritis and cancer progression. This model thus bridges fundamental autophagy biochemistry with translational gastric cancer pathology.
The ATG3 Knockout AGS Polyclonal Cells are ideally suited for a spectrum of functional studies using established autophagy assays. Autophagic flux can be measured via LC3-II western blotting with chloroquine or bafilomycin A1 blockade, complemented by immunofluorescence for LC3 puncta and electron microscopy. Cell viability (MTT/CCK-8), apoptosis (flow cytometry), and Transwell migration/invasion assays assess phenotypic consequences. Co-immunoprecipitation validates ATG3 interactions with LC3 or ATG7, while RT-qPCR quantifies autophagy gene expression. Applications extend to screening autophagy-modulating compounds, dissecting mTOR/AMPK signaling, and probing selective autophagy in infection models. For detailed product information, technical support, and ordering, please contact Ascent Research.
