The EIF2AK3 Knockout Ca Ski Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal population derived from the Ca Ski human cervical carcinoma cell line, with targeted disruption of the EIF2AK3 gene encoding PERK. This polyclonal format provides a heterogeneous mixture of knockout cells, avoiding clonal selection biases. The model is designed for investigating ER stress signaling and the unfolded protein response (UPR) in an epithelial cancer context.
Ca Ski is an adherent human epithelial cell line originating from a cervical epidermoid carcinoma, carrying integrated HPV-16 genomes and expressing viral oncoproteins E6 and E7. The cells are a well-established model for cervical cancer research, particularly for studies of HPV-mediated transformation, viral?Chost interactions, and therapeutic response.
EIF2AK3/PERK is an ER-resident kinase that senses ER stress through GRP78/BiP dissociation upon unfolded protein accumulation. Activated PERK phosphorylates eIF2??, attenuating global translation while selectively upregulating ATF4, which transcriptionally induces CHOP (DDIT3) and GADD34 (PPP1R15A). PERK signaling is triggered by oxidative stress, hypoxia, nutrient deprivation, and viral infection, and interfaces with apoptosis via TRAF2 and caspase-12. This pathway governs cell fate decisions between adaptation and programmed cell death under proteotoxic stress.
In the Ca Ski background, EIF2AK3 knockout allows dissection of PERK??s role in cervical carcinoma cells that naturally express HPV oncoproteins. The model is well-suited for examining how HPV-16 and PERK signaling co-regulate stress responses, chemoresistance, and apoptosis. It provides a platform to study synthetic lethality and identify vulnerabilities arising from loss of PERK function in an HPV-positive epithelial environment.
Research applications include quantitative analysis of UPR markers (phospho-eIF2??, ATF4, CHOP) by western blotting and RT-qPCR, and functional assays with ER stress inducers such as tunicamycin or thapsigargin. The cells can be used in ATF4-luciferase reporter systems, co-immunoprecipitation of PERK?CGRP78 complexes, immunofluorescence, and flow cytometry for apoptosis. They also support cell viability screens and studies on virus?Chost interactions. For further information, please contact Ascent Research.
