The EIF2AK2 Knockout K-562 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the K-562 cell line, featuring targeted disruption of the EIF2AK2 gene. This genetic perturbation abrogates expression of the double-stranded RNA (dsRNA)-activated protein kinase PKR, providing a loss-of-function model to dissect its roles in stress signaling, antiviral defense, and apoptosis. The polyclonal format ensures representation of diverse editing events across the population, making it suitable for pooled functional assays without clonal isolation.
K-562 cells are a suspension-adapted human chronic myelogenous leukemia (CML) cell line established from the pleural effusion of a 53-year-old female in blast crisis. They harbor the BCR-ABL fusion oncogene, which drives constitutive tyrosine kinase activity and leukemogenic signaling, and are widely used as a model for hematopoietic blast crisis CML. Their robust growth, ease of manipulation, and well-characterized signaling landscape make them an ideal host for gene knockout studies investigating oncogene-driven stress responses.
EIF2AK2 encodes PKR, a serine/threonine kinase activated by dsRNA, interferons (IFN-??/??/??), the dsRNA-binding protein PACT, tumor necrosis factor alpha (TNF??), lipopolysaccharide (LPS), and oxidative stress. Upon activation, PKR directly phosphorylates the ?? subunit of eukaryotic initiation factor 2 (eIF2??, encoded by EIF2S1), triggering global translation attenuation while selectively promoting translation of stress-responsive transcripts such as ATF4. Downstream, PKR engages IKK?? to activate NF-??B, stimulates JNK (MAPK8) signaling, and mediates apoptosis via p53 and caspase-8 pathways. PKR forms complexes with PACT, TRBP, TARBP2, viral dsRNA, and eIF2??, integrating innate immune and stress signaling cascades that ultimately regulate the integrated stress response through effectors like ATF4, DDIT3, PPP1R15A, and NFKB1.
In the BCR-ABL-driven K-562 background, PKR-mediated translational control and pro-apoptotic signaling intersect with oncogenic survival pathways, making this knockout model particularly valuable for studying CML biology. Disruption of EIF2AK2 enables dissection of how PKR influences BCR-ABL signal transduction, modulates sensitivity to tyrosine kinase inhibitors such as imatinib, and governs cell fate under proteotoxic or genotoxic stress. This system offers an isogenic platform to examine PKR-dependent and -independent contributions to leukemogenesis, drug resistance, and the antiviral state in a hematological malignancy context.
Researcher applications include western blot analysis of PKR and phosphorylated eIF2??, RT-qPCR profiling of interferon-stimulated genes (ISGs), flow cytometry-based apoptosis assays (Annexin V), NF-??B luciferase reporter assays, RNA-sequencing for transcriptome-wide stress responses, co-immunoprecipitation of PKR interactomes, viral replication kinetics, and drug sensitivity testing. For further information or to discuss custom requirements, please contact Ascent Research.
