The ATG7 Knockout 769-P Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human 769-P clear cell renal cell carcinoma (ccRCC) cell line. This population is engineered to disrupt the ATG7 gene, resulting in a loss-of-function model that abrogates ATG7-dependent autophagic activity. The polyclonal format provides a heterogeneous pool of edited alleles, offering a robust platform for studying autophagy disruption without the clonal selection bias inherent in single-cell-derived lines. This product is intended for advanced investigations into the role of autophagy in cancer biology, drug resistance, and metabolic signaling pathways.
The 769-P host cell line is a widely characterized model of ccRCC, originally established from a primary clear cell adenocarcinoma of the kidney. These cells harbor a biallelic loss-of-function mutation in the VHL tumor suppressor gene, leading to constitutive stabilization of hypoxia-inducible factors (HIF-1?? and HIF-2??). The resulting pseudo-hypoxic state drives a transcriptional program that promotes angiogenesis, metabolic reprogramming, and enhanced stress adaptation. The 769-P line is particularly relevant for dissecting the molecular mechanisms underlying renal carcinogenesis, VHL/HIF signaling, and mTOR pathway dysregulation, all of which intersect with the autophagy machinery.
ATG7 encodes a ubiquitin-like E1-activating enzyme essential for macroautophagy. It catalyzes the activation of ATG12 for conjugation to ATG5 and primes LC3/GABARAP proteins for lipidation, two ubiquitination-like cascades crucial for autophagosome elongation. ATG7 activity is regulated by mTORC1 suppression and AMPK activation, and its transcription is induced by FOXO, TFEB, MITF, and HIF-1?? under stress conditions. The enzyme functions in concert with ATG12, ATG5, ATG10, ATG3, and ATG16L1. Downstream, ATG7-dependent lipidation of LC3B enables autophagic cargo sequestration and degradation of p62/SQSTM1, a process vital for mitochondrial quality control.
In the 769-P ccRCC background, VHL loss stabilizes HIF-??, which transcriptionally upregulates ATG7. This places autophagy as a potential survival mechanism under the metabolic stress characteristic of renal tumors. ATG7 disruption in this background enables dissection of autophagy??s role in ccRCC cell survival, metabolic adaptation, and therapeutic resistance. This model is particularly valuable for testing whether autophagy inhibition enhances sensitivity to mTOR inhibitors, and for identifying ATG7-dependent vulnerabilities in clear cell carcinoma.
Typical applications include monitoring autophagy flux via LC3B Western blotting and p62/SQSTM1 accumulation, and fluorescence microscopy for LC3 puncta formation. Functional assays such as cell viability under nutrient deprivation, mitophagy flux analysis, and drug sensitivity profiling with mTOR inhibitors can identify autophagy-dependent phenotypes. These cells also serve as a crucial negative control for pharmacological autophagy modulators. For additional information, please contact Ascent Research.
