The EIF2AK2 Knockout HT29 Polyclonal Cells from Ascent Research are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma line. This product features targeted disruption of the EIF2AK2 gene, encoding protein kinase R (PKR), yielding a heterogeneous pool of edited cells for robust loss-of-function studies. The polyclonal format avoids clonal biases and provides a representative genetic knockout model for downstream functional assays.
HT29 cells are a well-characterized model of colorectal adenocarcinoma, carrying BRAF V600E and TP53 mutations while remaining microsatellite stable and competent for enterocytic differentiation. These features render the cells particularly relevant for investigating signaling crosstalk between oncogenic drivers and stress-response pathways in intestinal epithelial biology and colorectal cancer.
EIF2AK2 encodes the dsRNA-dependent serine/threonine kinase PKR, a pivotal sensor in antiviral innate immunity and the integrated stress response. PKR is activated by dsRNA, interferons (IFN-??/??/??), TNF??, and the activator PACT, while the inhibitor p58IPK suppresses its function. Active PKR phosphorylates eIF2??, attenuating global translation while selectively upregulating ATF4. PKR also promotes NF-??B activation via I??B phosphorylation and engages p53-dependent apoptosis. It interacts with proteins such as PACT, TRBP, TARBP2, NCK1, and HSP90, and operates downstream of interferon receptors (JAK1/TYK2) and pattern recognition receptors RIG-I and MDA5.
In the HT29 background, EIF2AK2 knockout enables detailed dissection of PKR??s role amid constitutive MAPK signaling from BRAF V600E and compromised p53 apoptosis. The model permits analysis of how PKR loss alters translational control, stress adaptation, and inflammatory signaling, impacting colorectal cancer cell phenotype, differentiation, and response to therapeutic stresses. It is also valuable for exploring PKR??s functions in intestinal epithelial homeostasis and inflammatory conditions.
Applications include quantitative Western blotting for phospho?eIF2?? and eIF2??, RT?qPCR for ATF4, CHOP, and interferon?stimulated genes, NF???B luciferase reporters, apoptosis flow cytometry, immunofluorescence, RNA?seq profiling, and co?immunoprecipitation of PKR complexes. Researchers can employ these cells to study antiviral innate immunity, translational regulation, cancer biology, integrated stress responses, and to screen for PKR modulators in drug discovery, with relevance to Alzheimer??s and diabetes. For detailed inquiries, contact Ascent Research.
