The EIF2AK2 Knockout HCT 116 Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal knockout cell population targeting the EIF2AK2 gene in human HCT 116 colorectal carcinoma epithelial cells. This pooled population harbors heterogeneous gene disruptions at the EIF2AK2 locus, providing a robust loss-of-function model for studying protein kinase R (PKR)-dependent signaling without clonal isolation artifacts.
The HCT 116 host cell line is derived from a human male colorectal adenocarcinoma and is widely used in cancer research due to its well-characterized genetic background. It carries an activating KRAS G13D mutation, retains wild-type TP53 status, and displays microsatellite instability-high (MSI-H) phenotype, making it particularly valuable for investigations into colorectal tumorigenesis, DNA mismatch repair deficiency, and targeted therapy responses.
EIF2AK2 encodes PKR, a dsRNA-activated kinase pivotal in innate antiviral immunity. Upon binding dsRNA or interacting with PACT/PRKRA, PKR autophosphorylates and then phosphorylates eIF2?? (EIF2S1), leading to global translation inhibition and selective expression of stress-responsive factors like ATF4 and CHOP. PKR signaling also engages NF-??B via IKK complex stimulation and can induce apoptosis through p53 and JNK. Interacting partners such as TRBP, Hsp90, and p58IPK modulate its activity, linking viral detection to cellular stress responses.
In the context of HCT 116 colorectal cancer cells, EIF2AK2 knockout provides a powerful tool to dissect the intersection between innate immune signaling and oncogenic pathways. Given the KRAS-driven and MSI-H background, this knockout model enables the study of how PKR activity influences tumor cell survival, chemosensitivity, and interactions with the tumor microenvironment. The dual role of PKR in promoting apoptosis and translational arrest may be particularly relevant in colorectal carcinomas that often exhibit chronic inflammatory signaling.
This EIF2AK2 knockout model enables multifaceted research, from antiviral innate immunity studies employing viral plaque reduction assays and ISG RT-qPCR to translational control investigations via phospho-eIF2?? Western blotting. Co-immunoprecipitation can map PKR protein interactions, while apoptosis pathways are probed using flow cytometry for Annexin V. Drug target evaluation is supported by MTT viability assays, and transcriptional outcomes can be assessed through RNA-seq or ISRE luciferase reporters. For further information or custom service inquiries, please contact Ascent Research.
