The EIF5A2 Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human liver adenocarcinoma cell line SK-HEP-1. This product comprises a heterogeneous pool of cells carrying targeted disruptions in the EIF5A2 gene, generated via CRISPR/Cas9-mediated gene disruption. It provides a versatile loss-of-function model for investigating oncogenic translation in hepatocellular carcinoma. The polyclonal format preserves population-level diversity, enabling robust phenotypic comparisons without the biases of single-cell clones.
SK-HEP-1 is an immortalized, epithelial cell line established from a liver adenocarcinoma patient. Widely employed in hepatic cancer research, these cells exhibit key features of hepatocellular carcinoma, including dysregulated proliferation and apoptosis resistance. Their genetic tractability and stable growth characteristics make them a favored host for CRISPR-based gene editing. When modified with CRISPR/Cas9, SK-HEP-1 cells retain their malignant properties while permitting dissection of individual gene contributions to liver cancer pathology.
EIF5A2 encodes eukaryotic translation initiation factor 5A2, a specialized elongation factor that facilitates synthesis of proteins containing consecutive proline motifs. Its activity requires post-translational hypusination by deoxyhypusine synthase (DHPS), a process modulated by polyamine metabolism and mTOR signaling. EIF5A2 predominantly functions on the 60S ribosomal subunit to resolve ribosome stalling at polyproline sequences. Upstream, EIF5A2 expression is driven by MYC transcriptionally and controlled translationally by the mTOR/4E-BP1 axis. Spermidine, a polyamine substrate for DHPS, further regulates hypusination. Consequently, EIF5A2 integrates proliferative and survival signals to enhance translation of oncoproteins such as cyclin D1 and Bcl-2. Knockout of EIF5A2 disrupts this output, impairing cell cycle progression and survival pathways.
In the SK-HEP-1 hepatocellular carcinoma context, loss of EIF5A2 likely attenuates translation of key mediators of tumor aggressiveness, including cyclin D1 and Bcl-2, thereby reducing proliferation and sensitizing cells to apoptosis. Because SK-HEP-1 retains functional p53, the knockout may further compromise growth under stress. The polyclonal knockout population minimizes clonal artifacts and better reflects heterogeneous tumor cell behavior, making it an ideal model for studying translational control mechanisms in liver cancer.
Applications include MTT-based proliferation assays, flow cytometric apoptosis detection, polysome profiling for translational efficiency, and western blotting for downstream effectors (cyclin D1, Bcl-2, p53). Migration and invasion can be assessed by wound-healing or transwell assays, with molecular validation via RT-qPCR. These cells are also suitable for drug sensitivity screens to identify agents whose activity is modulated by EIF5A2 status, and for epistasis studies with mTOR or polyamine pathway modulators. For further information or ordering, please contact Ascent Research.
