The EIF5A2 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HEK293T human embryonic kidney cell line, designed to disrupt the EIF5A2 gene. This loss-of-function model enables investigation of eukaryotic translation initiation factor 5A2 (EIF5A2), a critical elongation factor that resolves ribosome stalling at polyproline motifs. The polyclonal nature preserves genetic heterogeneity, avoiding clonal selection artifacts, and provides a powerful tool for studying EIF5A2-dependent processes in an easily transfectable epithelial context.
The parental HEK293T line is a widely utilized derivative of the HEK293 cell line, stably expressing the SV40 large T-antigen. This modification enables high-level episomal replication of plasmids containing the SV40 origin, leading to exceptional transfection efficiency and robust recombinant protein expression. HEK293T cells are of human embryonic kidney origin, exhibiting epithelial morphology and providing a well-characterized model for studying signal transduction, translation regulation, and cancer-relevant pathways. Their non-tumorigenic yet experimentally permissive nature suits dissection of oncogenic mechanisms.
EIF5A2 functions as a translation elongation factor that, upon hypusination, promotes efficient translation of polyproline-containing proteins. Hypusination is sequentially catalyzed by deoxyhypusine synthase (DHPS) and deoxyhypusine hydroxylase (DOHH), using spermidine as a substrate. EIF5A2 activity is integrated into critical signaling networks: it is transcriptionally activated by MYC and HIF1A, and its hypusination status is responsive to mTORC1 and polyamine metabolism. Active, hypusinated EIF5A2 facilitates translation of key downstream targets such as cyclin D1, driving cell cycle progression, and the transcription factor Snail, a master regulator of epithelial-mesenchymal transition (EMT). Additionally, EIF5A2 interacts directly with ribosomal subunits and exportin-1 (XPO1) for proper localization, and influences apoptosis by modulating BCL-2 family protein expression, thereby linking translational control to cell survival pathways.
In the HEK293T background, disruption of EIF5A2 provides a controlled system to delineate its function in translation elongation and associated signaling cascades. Given the kidney epithelial origin, this model is particularly relevant for studying the molecular basis of EMT and cell proliferation??processes frequently dysregulated in carcinomas. Overexpression of EIF5A2 has been implicated in hepatocellular, colorectal, and ovarian cancers, as well as glioblastoma, making this knockout tool invaluable for dissecting oncogenic translation programs. By removing EIF5A2 function, researchers can examine the dependency of polyproline-containing gene expression on the hypusination pathway and evaluate the consequent effects on cell growth, migration, and apoptotic signaling within an epithelial context.
This knockout cell product enables investigation of translation elongation control, including polysome profiling and puromycin incorporation. Western blotting verifies EIF5A2 and hypusine loss; RT-qPCR and luciferase reporter systems quantify target mRNA translation. Colony formation, wound healing, and transwell assays measure proliferation and migration. These cells aid identification of EIF5A2-dependent mRNAs. For further information, contact Ascent Research.
