This product is a CRISPR/Cas9-edited polyclonal knockout cell population of the NCI-H1975 human non-small cell lung adenocarcinoma line with targeted EIF4A2 gene disruption. The polyclonal pool captures heterogeneous edited alleles, mirroring native tumor genetic variability without clonal selection pressures. EIF4A2 encodes an ATP-dependent RNA helicase essential for unwinding 5′ UTR structures during cap-dependent translation initiation, and its knockout provides a loss-of-function model for studying translational control in lung cancer.
NCI-H1975 is an epithelial cell line from pleural effusion of a lung adenocarcinoma patient, harboring EGFR L858R and T790M mutations and p53 deficiency, representing a clinically relevant model for tyrosine kinase inhibitor resistance and advanced NSCLC. It is widely used to study oncogenic signaling, apoptosis, and drug response, particularly to mTOR and translation inhibitors. The cell line’s reliance on cap-dependent translation for pro-survival factor synthesis underscores the value of this EIF4A2 knockout for dissecting tumor biology.
EIF4A2 functions as a core component of the eIF4F complex, interacting with eIF4E, eIF4G, and PDCD4. Its helicase activity is stimulated by eIF4B and controlled by mTORC1 and MAPK/ERK via MNK kinases. This integrates growth factor signals (EGF, insulin), nutrient status, and stress (UPR, hypoxia) to drive selective translation of oncogenic mRNAs such as MYC, BCL-2, cyclin D1, VEGF, and survivin, thereby regulating proliferation, survival, and angiogenesis.
Loss of EIF4A2 in the NCI-H1975 background is predicted to impair cap-dependent translation, particularly of mRNAs with structured 5′ UTRs driving malignancy. With EGFR mutations and p53 loss, EIF4A2 knockout may uncover synthetic vulnerabilities and alter sensitivity to mTORC1 or eIF4A inhibitors (rapamycin, silvestrol). This model offers a platform to explore the crosstalk between kinase signaling and translation initiation and to identify translational addiction mechanisms in lung adenocarcinoma.
Researchers can apply this polyclonal knockout in polysome profiling, cap-dependent translation reporter assays, and co-immunoprecipitation to monitor eIF4F complex. Additional assays include western blotting, RT-qPCR, viability and apoptosis tests, colony formation, and migration studies. The model is ideal for drug sensitivity screens with translation inhibitors (silvestrol, rapamycin) and for RNA-seq or ribosome profiling. By dissecting EIF4A2-dependent translation networks, this product supports target validation, resistance research, and functional genomics in NSCLC. Contact Ascent Research for technical details.
