The EIF2AK3 Knockout HGC-27 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population in which the EIF2AK3 gene (encoding PERK) has been disrupted. This loss-of-function model is generated in the HGC-27 gastric carcinoma cell line and provides a heterogeneous population of edited cells, each carrying CRISPR-mediated gene edits that abrogate functional PERK expression. The polyclonal nature ensures representation of diverse editing outcomes while enabling pooled functional studies without clonal artifacts. This product is designed for researchers investigating endoplasmic reticulum (ER) stress signaling, the unfolded protein response (UPR), and the integrated stress response in the context of gastric cancer biology.
The HGC-27 host cell line is of human epithelial origin, originally established from a metastatic lymph node of a gastric adenocarcinoma. These cells are widely used as a model system for gastric cancer, particularly for studying mechanisms of lymph node metastasis, invasive growth, and therapeutic resistance. HGC-27 cells retain key characteristics of gastric carcinoma, including their epithelial morphology and responsiveness to ER stress triggers, making them an appropriate background in which to examine the tumor-relevant functions of PERK. The knockout of EIF2AK3 in this metastatic setting offers a precise genetic tool for dissecting PERK-dependent signaling pathways in a disease-relevant cellular context.
EIF2AK3 encodes the endoplasmic reticulum stress sensor PERK, a type I transmembrane kinase that plays a central role in the UPR. Under basal conditions, PERK is maintained in an inactive state through binding to the chaperone GRP78/BiP. Upon accumulation of misfolded proteins within the ER, as well as during hypoxia, nutrient deprivation, or oxidative stress, GRP78 dissociates, allowing PERK dimerization and autophosphorylation. Activated PERK directly phosphorylates the alpha subunit of eukaryotic initiation factor 2 (eIF2??) on serine 51, which attenuates global cap-dependent translation while selectively enhancing the translation of the transcription factor ATF4. ATF4 subsequently upregulates downstream targets including CHOP (DDIT3), GADD34, and autophagy-related proteins, driving adaptive pro-survival or pro-apoptotic programs. PERK also interacts with TRAF2, IRE1, and the inhibitory protein p58IPK, integrating diverse signals to modulate cell fate decisions. Disruption of EIF2AK3 thus cripples this critical pathway, blocking eIF2?? phosphorylation and downstream UPR transcriptional responses.
The EIF2AK3 knockout in HGC-27 cells holds particular significance for gastric cancer research, as PERK-mediated UPR activation is frequently co-opted by tumors to survive the hostile microenvironment of metastatic sites. HGC-27 cells, being metastatic in origin, likely rely on PERK signaling to cope with proteotoxic stress, hypoxia, and nutrient scarcity encountered during dissemination and colonization. By eliminating PERK function, this polyclonal knockout model enables direct interrogation of whether the UPR contributes to metastatic cell viability, migration, invasion, and resistance to chemotherapeutic agents. Researchers can use this system to delineate the role of PERK in balancing autophagy and apoptosis downstream of ER stress, and to evaluate therapeutic strategies targeting PERK in advanced gastric carcinoma.
This knockout product is suited for a broad range of experimental applications. Users can assess PERK activity through Western blot detection of phospho-eIF2?? and monitor downstream transcriptional changes via RT-qPCR for ATF4 and CHOP. Transcriptome-wide effects can be captured by RNA-seq, while functional outcomes such as apoptosis can be quantified by flow cytometry. Reporter assays using ATF4-luciferase constructs allow real-time monitoring of the integrated stress response. Additional techniques include immunofluorescence for ER markers, co-immunoprecipitation of PERK with GRP78 to study complex dynamics, and drug sensitivity assays with PERK inhibitors to explore pharmacological interventions. Migration and invasion assays further extend the model’s utility in metastasis research. For further technical details and custom inquiries, please contact Ascent Research.
