The EIF2AK3 Knockout NCI-H1299 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in which the EIF2AK3 gene has been disrupted, abolishing functional PERK expression. This polyclonal format provides a heterogeneous loss-of-function model that is suitable for studying PERK-dependent signaling without selection of a single clone, preserving population-level diversity. The product is designed for researchers investigating endoplasmic reticulum (ER) stress and the unfolded protein response (UPR) in a human lung adenocarcinoma background.
The host cell line, NCI-H1299, was derived from a lymph node metastasis of a lung adenocarcinoma from a 53-year-old male. It is a widely utilized model for non-small cell lung cancer (NSCLC) metastasis, characterized by its aggressive behavior and ability to form tumors in vivo. NCI-H1299 cells lack expression of p53 protein, which contributes to their genetic instability and is relevant for apoptosis studies. Their metastatic origin makes them particularly valuable for examining stress-adaptive mechanisms that support cancer cell survival and dissemination.
EIF2AK3 encodes the ER-resident kinase PERK, a central sensor of ER stress. Upon accumulation of misfolded proteins, GRP78/BiP dissociates from PERK, triggering its activation and subsequent phosphorylation of the ?? subunit of eukaryotic initiation factor 2 (eIF2??). This attenuates global translation while selectively upregulating ATF4, which induces transcription of CHOP (DDIT3) and GADD34, driving either adaptive or apoptotic programs. PERK also directly phosphorylates NRF2, promoting its nuclear translocation and antioxidant gene expression. Negative regulation is mediated by P58IPK and the GADD34?CPP1 phosphatase complex that dephosphorylates eIF2??. Downstream effectors include the pro-apoptotic BAX/BAK pathway and autophagy components such as Beclin-1 and LC3.
In NCI-H1299 cells, knockout of EIF2AK3 abrogates PERK-mediated eIF2?? and NRF2 phosphorylation, disrupting the integrated stress response (ISR) and weakening the cell??s ability to manage proteotoxic stress. This renders the cells more susceptible to ER stress-inducing agents such as tunicamycin, thapsigargin, and bortezomib, while altering the balance between survival, apoptosis, and autophagy. The model therefore provides a physiologically relevant platform to dissect the role of PERK signaling in NSCLC metastasis, where ER stress adaptation is often linked to drug resistance and tumor progression. It also enables studies of PERK-dependent translational control and antioxidant defense in a cancer context.
The EIF2AK3 Knockout NCI-H1299 Polyclonal Cells are applicable across a range of functional assays, including Western blotting for phosphorylated eIF2??, ATF4, and CHOP, as well as RT-qPCR analysis of UPR target genes. They can be employed in drug sensitivity screening for cisplatin, bortezomib, and PERK inhibitors, and in flow cytometry-based apoptosis assays using Annexin V/PI staining. Additional uses include autophagy flux analysis, polysome profiling to assess translation, and transcriptomic studies via RNA-seq. For further details or to request a quote, please contact Ascent Research.
