The EIF2AK3 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population targeting EIF2AK3 in HEK293T cells. This heterogeneous pool of loss-of-function cells avoids clonal artifacts, providing a robust model without single-cell cloning. The polyclonal format captures diverse gene-disruption events, enabling population-based functional studies.
HEK293T cells are immortalized human embryonic kidney epithelia transformed with sheared adenovirus type 5 DNA and expressing SV40 large T antigen. These features ensure high transfectability and robust protein production, making them a mainstay for transient/stable expression, lentiviral generation, and pathway analysis. Their epithelial origin and vigorous growth suit them for studying stress-responsive signaling and gene regulation.
EIF2AK3 encodes PERK, an ER-resident kinase acting as a primary sensor of the unfolded protein response. ER stress triggers BiP/GRP78 dissociation, PERK oligomerization, and autophosphorylation. Active PERK phosphorylates eIF2??, transiently attenuating global translation while selectively upregulating ATF4 translation. ATF4 induces stress-responsive genes including CHOP, GADD34, and NRF2-dependent antioxidants. This PERK-eIF2??-ATF4 signaling branch integrates with the integrated stress response, governing cell fate decisions between adaptation and apoptosis.
In HEK293T cells, EIF2AK3 knockout removes the principal ER stress-sensing kinase, enabling dissection of PERK-dependent and -independent UPR arms. Researchers can examine how loss of eIF2?? phosphorylation influences translational regulation, ATF4/CHOP induction, and apoptosis upon stressors like tunicamycin or thapsigargin. This model is valuable for exploring PERK roles in cancer drug resistance, metabolic adaptation, and ER stress-linked neurodegeneration. The polyclonal population minimizes clonal bias and captures diverse knockout phenotypes, enhancing reproducibility.
Applications include PERK-eIF2??-ATF4 pathway analysis, PERK inhibitor screening, modeling of Wolcott-Rallison syndrome ER stress, and studies of ER stress in cancer and neurodegeneration. Common assays involve Western blotting for phospho-eIF2?? and PERK, RT-qPCR for ATF4/CHOP/GADD34, apoptosis assays with caspase or annexin V, and cell viability tests. For more information, please contact Ascent Research.
