The EIF2AK1 Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the EIF2AK1 gene in the human SK-HEP-1 cell line. This pool of edited cells provides a loss-of-function model for studying EIF2AK1 (heme-regulated inhibitor, HRI) function.
SK-HEP-1 is a human liver adenocarcinoma cell line derived from ascitic fluid, exhibiting adherent, epithelial morphology with endothelial characteristics. It serves as a model for hepatocellular carcinoma and endothelial biology, including angiogenesis studies.
EIF2AK1 encodes a stress-regulated kinase activated by heme deficiency, oxidative stress, heat shock, and proteasome inhibition. Upon activation, it phosphorylates eIF2??, attenuating global translation while selectively enhancing ATF4 translation. ATF4 induces stress-responsive genes such as CHOP and GADD34 (PPP1R15A). EIF2AK1 interacts with heme, HSP90, and CDC37; heme binding inhibits kinase activity, while its dissociation triggers signaling. The downstream targets phospho-eIF2??, ATF4, CHOP, and GADD34 establish a negative feedback loop via PPP1R15A-mediated eIF2?? dephosphorylation, restoring homeostasis.
In the SK-HEP-1 context, EIF2AK1 knockout enables dissection of heme-regulated translational control in a liver cancer model. This is relevant for studying tumor cell adaptation to stress, drug resistance, and the integrated stress response’s role in hepatocellular carcinoma progression.
Applications include western blotting for phospho-eIF2?? and ATF4, RT-qPCR for CHOP and GADD34, cell viability assays under stress, polysome profiling, and flow cytometry for apoptosis. The polyclonal population supports large-scale experiments, maintaining genetic heterogeneity beneficial for population-based stress response studies. For further details, contact Ascent Research.
