The EIF2AK3 Knockout Huh-7 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the EIF2AK3 gene, which encodes the ER stress sensor kinase PERK. This product was generated by introducing targeted gene disruptions in Huh-7 cells via CRISPR/Cas9, producing a heterogeneous pool of cells with disrupted EIF2AK3 alleles. The polyclonal format provides a robust model for studying PERK-dependent signaling without clonal selection bias. This pool is suitable for functional assays examining the unfolded protein response (UPR) and integrated stress response (ISR) pathways.
The host cell line, Huh-7, is a well-differentiated hepatocellular carcinoma from a male Japanese patient. These liver epithelial cells retain hepatocyte characteristics, including metabolic activity and liver-specific markers, making them relevant for hepatic studies. Huh-7 cells are widely used in cancer biology, virology, and drug metabolism research. Their epithelial origin provides a context for investigating stress signaling that impacts hepatocyte survival and transformation.
EIF2AK3 (PERK) is a type I transmembrane protein kinase in the ER membrane, serving as a primary ER stress sensor. Upon misfolded protein accumulation, PERK activation occurs through dissociation from BiP/GRP78. Activated PERK phosphorylates eIF2??, causing global translational attenuation while allowing selective ATF4 translation. ATF4 promotes adaptive genes and, under unresolved stress, induces the pro-apoptotic factor CHOP (DDIT3). PERK signaling intersects with IRE1 and ATF6 pathways, modulated by P58IPK/DNAJC3, calreticulin, and PDI. Reactive oxygen species and calcium depletion further regulate the pathway, converging on downstream effectors including GADD34, NRF2, and NF-??B.
In Huh-7 hepatocellular carcinoma cells, PERK-mediated UPR signaling adapts to the stressful tumor microenvironment of hypoxia, nutrient deprivation, and oxidative stress. EIF2AK3 knockout disrupts this adaptation, sensitizing cells to apoptotic stimuli and altering autophagy and metabolism. This model is valuable for dissecting PERK’s roles in liver cancer cell survival, drug resistance, and the pro-survival/pro-death UPR balance. Given the liver’s high secretory load, these cells provide a platform for studying ER stress-related hepatic pathologies, including steatosis and hepatocellular carcinoma progression.
Researchers can use these EIF2AK3 knockout Huh-7 polyclonal cells in diverse workflows. Applications include characterizing PERK-dependent eIF2?? phosphorylation via western blotting, assessing UPR target genes (e.g., ATF4, CHOP) by RT-qPCR, and evaluating responses to ER stress inducers like tunicamycin or thapsigargin. The model supports cell viability, apoptosis, ATF4-luciferase reporter assays, phospho-signaling analysis, and immunofluorescence for ER stress markers. These cells are also suited for drug discovery targeting the PERK pathway, metabolic disease research, and neurodegeneration studies. For further information, please contact Ascent Research.
