The EIF2AK4 Knockout A2780 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the human ovarian adenocarcinoma A2780 cell line, designed for constitutive disruption of the EIF2AK4 gene. This loss-of-function model enables investigation of GCN2-dependent stress signaling without clonal selection artifacts.
A2780 cells, established from an untreated patient with ovarian endometrioid adenocarcinoma, exhibit adherent epithelial morphology and are inherently cisplatin-sensitive. They serve as a standard model for ovarian cancer drug response and chemoresistance studies.
EIF2AK4 (GCN2) is a serine/threonine kinase that functions as a primary sensor of amino acid deprivation. Activated by uncharged tRNAs during amino acid scarcity, GCN2 phosphorylates eIF2??, leading to global translation attenuation and selective translation of ATF4, the master transcription factor of the integrated stress response (ISR). GCN2 interacts with GCN1 and GCN20 at the ribosome and is modulated by IMPACT. The GCN2-eIF2??-ATF4 axis induces expression of stress-adaptive genes including CHOP, GADD34, ASNS, REDD1, and ATF3, which regulate amino acid metabolism, autophagy, and apoptosis. Additional activation stimuli include UV irradiation, oxidative stress, and mTORC1 inhibition, highlighting cross-talk with nutrient-sensing pathways such as mTORC1 via Sestrin2.
In A2780 cells, the ISR is crucial for survival under nutrient stress and chemotherapeutic challenge. EIF2AK4 knockout disrupts amino acid sensing and the downstream phosphorylation of eIF2??, impairing ATF4-mediated adaptive responses. This may sensitize cells to cisplatin and paclitaxel, making the model valuable for exploring GCN2??s role in chemoresistance, metabolic reprogramming, and autophagy. Additionally, it facilitates identification of synthetic lethal targets and evaluation of ISR inhibitors.
Applications include western blotting for phospho-eIF2?? and ATF4, RT-qPCR of ISR target genes, viability assays under amino acid starvation, colony formation, and apoptosis analysis. Drug sensitivity screening with cisplatin or paclitaxel, migration/invasion assays, RNA-seq, and polysome profiling are also compatible. For additional information, please contact Ascent Research.
