The EIF2AK3 Knockout K-562 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the EIF2AK3 gene in the human K-562 cell line. This engineered model provides a loss-of-function system to study the ER stress sensor kinase PERK. The polyclonal format comprises a heterogeneous pool of cells with diverse gene-disruption events at the target locus, enabling population-level analysis of knockout effects without single-cell cloning.
K-562 is a chronic myelogenous leukemia cell line established from the pleural effusion of a patient in blast crisis. It is a widely used hematopoietic progenitor model with the capacity to differentiate along erythroid and megakaryocytic lineages. This suspension cell line is readily transfectable, making it amenable to CRISPR/Cas9-mediated gene editing for investigating signaling pathways relevant to leukemia and hematopoiesis.
EIF2AK3 encodes the ER-resident kinase PERK, a central mediator of the unfolded protein response. Under ER stress, PERK is activated by dissociation from BiP/GRP78, leading to dimerization, autophosphorylation, and subsequent phosphorylation of eIF2?? at Ser51. This inhibits global cap-dependent translation while selectively increasing translation of ATF4, a transcription factor that induces downstream targets such as CHOP, GADD34, and TRB3. PERK signaling is modulated by upstream stressors including hypoxia and nutrient deprivation, and it interacts with IRE1??, ATF6, and TRAF2, integrating UPR signals to regulate redox homeostasis, autophagy, and apoptosis.
In K-562 leukemia cells, the UPR supports survival under oncogenic and therapeutic stress. Disruption of EIF2AK3 enables researchers to dissect PERK-dependent versus -independent ER stress responses, particularly in contexts such as proteasome inhibitor resistance and hypoxia adaptation. This knockout model is relevant for studying the molecular basis of Wolcott-Rallison syndrome, diabetes, skeletal dysplasia, cancer progression, and neurodegenerative diseases where PERK signaling is implicated.
These polyclonal knockout cells are suitable for a range of experimental applications, including Western blot analysis of p-eIF2??, ATF4, and CHOP following ER stress induction with thapsigargin or tunicamycin. They support RT-qPCR profiling of UPR target genes, ATF4 luciferase reporter assays, and RNA-seq transcriptomic studies. Viability assays (MTT, CellTiter-Glo) and flow cytometry for apoptosis (Annexin V) allow functional evaluation of stress responses, making the model valuable for PERK inhibitor screening and metabolic disorder research. For further technical information, please contact Ascent Research.
