The EIF5A2 Knockout NCI-H1975 Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal population with targeted disruption of the EIF5A2 locus in the NCI-H1975 human lung adenocarcinoma line. As a polyclonal knockout cell population, it provides a genetically heterogeneous loss-of-function model that avoids clonal selection biases while abrogating functional EIF5A2 protein expression. This reagent is optimized for investigating the downstream consequences of impaired EIF5A2-dependent translation elongation.
The parental NCI-H1975 line derives from a 62-year-old female with non-small cell lung adenocarcinoma and carries EGFR L858R/T790M mutations, which result in constitutive kinase activity and acquired resistance to first-generation EGFR inhibitors. These cells maintain epithelial morphology and activated signaling through mTORC1 downstream of EGFR, making them a clinically relevant model for studying targeted therapy resistance and metastatic progression.
EIF5A2 is a translation elongation factor that selectively enables synthesis of proteins containing polyproline stretches and stress-responsive regulators. Its activity requires post-translational hypusination by deoxyhypusine synthase (DHPS) and deoxyhypusine hydroxylase (DOHH). Transcriptionally, EIF5A2 is upregulated by MYC in response to mTORC1 and EGF/EGFR signaling, with regulatory inputs from TP53 and HIF1A. Hypusinated EIF5A2 interacts with the ribosome and eEF2 to facilitate translation of downstream targets including MYC, cyclin D1, MMPs, BCL2, and EMT transcription factors SNAI1 and ZEB1, thereby promoting proliferation, survival, invasion, and metastasis.
In the NCI-H1975 context, CRISPR-mediated disruption of EIF5A2 creates a powerful platform for dissecting translation-dependent mechanisms that sustain EGFR T790M/L858R-driven tumorigenicity and drug resistance. Because mutant EGFR signaling converges on mTORC1 and MYC to drive EIF5A2 transcription, knockout of this factor selectively ablates the translation of pro-oncogenic and pro-metastatic effectors, rendering cells vulnerable to apoptotic and anti-migratory consequences.
The EIF5A2 Knockout NCI-H1975 Polyclonal Cells support applications in cancer cell biology, translation control, drug resistance, and metastasis research. Common assays include Western blotting for EIF5A2 and hypusination, proliferation and transwell migration assays, apoptosis measurement by Annexin V/PI, RT-qPCR for downstream targets, polysome profiling, and ribosomal co-immunoprecipitation. This model enables detailed interrogation of mTORC1/MYC/EIF5A2 signaling and its role in lung adenocarcinoma. For further information, please contact Ascent Research.
