The ATG3 Knockout 769-P Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal cell population derived from the 769-P human clear cell renal cell carcinoma line. This product provides researchers with a heterogeneous pool of cells harboring targeted disruptions in the ATG3 gene, facilitating loss-of-function investigations without the confounding effects of clonal selection. Polyclonal populations are advantageous for collectively assessing gene function across diverse genetic edits, closely mimicking the heterogeneous nature of tumor biology.
The 769-P cell line was established from a primary clear cell renal cell carcinoma (ccRCC) and is widely utilized as an epithelial tumor model. These cells exhibit key hallmarks of ccRCC, including dysregulated mTOR pathway activity, hypoxia inducible factor stabilization, and altered autophagic responses. Their adherent growth and well-characterized signaling networks make them an ideal platform for studying autophagy-related mechanisms in kidney cancer, particularly in the context of metabolic adaptation and targeted therapy resistance.
ATG3 encodes an E2-like enzyme that catalyzes the conjugation of phosphatidylethanolamine (PE) to the ATG8 family of ubiquitin-like proteins, encompassing the LC3 (MAP1LC3A, MAP1LC3B) and GABARAP (GABARAP, GABARAPL1) subfamilies. This lipidation event is essential for autophagosome membrane elongation and cargo recognition. ATG3 activation is driven by the E1 enzyme ATG7 and the E3-like ATG5-ATG12 conjugate, operating downstream of the ULK1 complex and the class III PI3K complex. Nutrient status signals through mTOR kinase regulate ATG3 activity via phosphorylation of upstream initiation factors. ATG3 directly interacts with ATG7, ATG5, and ATG8 proteins to execute the conjugation cascade, underscoring its pivotal role in the autophagy core machinery.
Disruption of ATG3 in the 769-P renal carcinoma background abrogates ATG8 lipidation, offering a powerful tool to dissect autophagy-dependent processes in ccRCC. These cells enable interrogation of how loss of autophagic flux impacts tumor cell proliferation, survival under nutrient deprivation, and sensitivity to chemotherapeutic agents. Given the importance of autophagy in renal cancer drug resistance, this model provides a relevant system for exploring synthetic lethal interactions or identifying novel vulnerabilities that can be targeted therapeutically.
Typical applications for this polyclonal knockout model include autophagy flux analyses via immunoblotting for LC3-II conversion, fluorescence microscopy to monitor autophagosome accumulation, and flow cytometry using autophagy-sensitive probes. Co-immunoprecipitation of ATG3 with ATG7 or ATG8 proteins, along with RT-qPCR to verify gene disruption, allows detailed mechanistic studies. These cells are ideal for functional genomics screens, drug sensitization assays, and investigation of mTOR-dependent signaling pathways. For additional product details and technical support, please contact Ascent Research.
