The EIF2AK3 Knockout HeLa Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal population with disruption of the EIF2AK3 gene, which encodes the ER stress sensor PERK. This loss-of-function model is designed for investigating ER stress signaling and the integrated stress response in a human epithelial context. The polyclonal format avoids clonal selection artifacts, providing a more representative knockout model for functional studies.
HeLa cells are an immortalized epithelial line derived from a cervical adenocarcinoma, widely used in cancer biology, signal transduction, and drug discovery. Their robust growth and well-characterized genetic landscape facilitate reproducible experiments. HeLa cells exhibit canonical UPR activation upon ER stress, making them a suitable host for interrogating PERK-dependent pathways.
EIF2AK3/PERK is activated by ER stress, such as unfolded proteins or Ca2? depletion, which triggers dissociation of the inhibitory chaperone BiP/GRP78. Activated PERK phosphorylates eIF2??, globally suppressing translation while selectively increasing ATF4 synthesis. ATF4 transcriptionally induces CHOP and GADD34, which mediate adaptive or apoptotic outcomes. Additionally, PERK phosphorylates NRF2, linking the UPR to antioxidant responses. PERK functions in concert with IRE1?? and ATF6, integrating stress signals to determine cell fate. Key upstream regulators include BiP and PDIA6; downstream effectors encompass the transcription factors ATF4 and CHOP.
In the HeLa cancer cell background, PERK signaling often promotes survival under chronic ER stress, a hallmark of solid tumors. Knockout of EIF2AK3 allows researchers to dissect the pro-survival and pro-death functions of PERK, and to evaluate its contribution to drug resistance and metabolic adaptation. This model is particularly relevant for studying the PERK-eIF2??-ATF4-CHOP axis in cancer biology and for identifying vulnerabilities that can be exploited therapeutically.
Typical applications include ER stress induction with tunicamycin or thapsigargin followed by Western blotting for phospho-eIF2??, ATF4, and CHOP, as well as RT-qPCR of UPR target genes. The knockout cells are also suitable for apoptosis assays, immunofluorescence localization of PERK, reporter assays, and RNA-seq profiling of stress-induced transcriptomes. Researchers investigating Wolcott-Rallison syndrome, diabetes, neurodegeneration, or cancer drug resistance will find this model valuable. For further information, please contact Ascent Research.
