The EIF2AK2 Knockout 769-P Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the EIF2AK2 gene in the 769-P human renal epithelial cell line. This heterogeneous knockout pool, generated by CRISPR/Cas9-mediated gene disruption, provides a robust loss-of-function model for investigating EIF2AK2 (PKR) signaling without clonal selection biases. The polyclonal nature mimics population-level genetic variation often more relevant to tissue-level responses.
The 769-P cell line originates from a clear cell renal cell carcinoma and serves as a model for renal epithelial cells involved in absorptive and secretory functions. These adherent cells retain morphological and functional characteristics of proximal tubule epithelium, making them suitable for studies in renal cancer biology, drug transport, and signal transduction pathways relevant to kidney physiology and malignancy.
EIF2AK2 encodes protein kinase R (PKR), a dsRNA-activated serine/threonine kinase central to antiviral innate immunity and cellular stress responses. Upon stimulation by dsRNA, type I interferons, or the protein activator PACT (PRKRA), PKR autophosphorylates and directly phosphorylates eIF2?? at Ser51. This phosphorylation attenuates cap-dependent translation while selectively promoting translation of stress-responsive transcripts such as ATF4 and CHOP. Downstream of eIF2??, PKR signaling engages NF-kB and p38 MAPK cascades, leading to transcriptional induction of pro-inflammatory cytokines and apoptosis regulators. PKR physically interacts with TARBP2, TRBP, and RAX, and functions in complexes that integrate signals from pattern recognition receptors including TLR3, RIG-I, and MDA5.
In the 769-P renal carcinoma context, disruption of EIF2AK2 allows dissection of the kinase’s contributions to cancer cell survival, inflammatory signaling, and antiviral responses. Because clear cell renal cell carcinoma frequently exhibits altered interferon and stress signaling, this knockout model is particularly valuable for examining how PKR affects chemosensitivity, apoptosis resistance, and the balance between translational control and NF-kB?Cdriven transcription. The model also enables evaluation of downstream targets such as p53, STAT1, and Bcl-2 family members in kidney cancer.
This polyclonal knockout product supports a wide range of downstream assays, including Western blotting for phospho-eIF2?? and CHOP, RT-qPCR analysis of interferon-stimulated genes, immunofluorescence localization of PKR and its interactors, flow cytometry for annexin V?Cbased apoptosis detection, and NF-kB luciferase reporter measurements. Additional applications encompass co-immunoprecipitation of PKR with PACT or eIF2??, drug sensitivity profiling, phospho-signaling arrays, and transcriptomic studies via RNA-seq. For further technical details or custom inquiries, please contact Ascent Research.
