The GPX7 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population generated by disrupting the GPX7 gene in the HAP1 human near-haploid cell line. This product provides a genetically heterogeneous pool of cells carrying targeted GPX7-inactivating modifications, creating a functional loss-of-function model for population-based assays. The polyclonal format minimizes clonal selection biases and ensures robust representation of diverse editing outcomes.
HAP1 cells, originating from a chronic myeloid leukemia patient, exhibit an adherent fibroblast-like morphology and possess a quasi-haploid karyotype that reduces genetic redundancy and simplifies knockout characterization. This feature, combined with their facile genetic manipulation, has established HAP1 cells as a preferred system for functional genomics screens, drug target validation, and systematic pathway interrogation in cancer and oxidative stress research.
GPX7 functions as an endoplasmic reticulum-resident glutathione peroxidase that reduces hydrogen peroxide and organic hydroperoxides, thereby preserving ER redox homeostasis. Its expression is transcriptionally controlled by NRF2 (NFE2L2) downstream of KEAP1, and is further induced by ATF4 and the ER stress sensors PERK and IRE1?? during unfolded protein response (UPR) activation. GPX7 forms functional complexes with GRP78/BiP, ERO1L, and PDI family oxidoreductases, integrating oxidative stress sensing with protein folding machinery. Loss of GPX7 culminates in dysregulated intracellular ROS, amplified UPR signaling, and enhanced susceptibility to CHOP-mediated apoptosis, while also perturbing NRF2-driven antioxidant genes such as GCLC, GCLM, and HMOX1.
Knockout of GPX7 in HAP1 cells abolishes a critical ER peroxide scavenger, causing elevated ROS levels that potentiate oxidative stress and UPR-mediated cell death. This sensitization provides a cell-based model to study ER redox imbalance in diseases where GPX7 dysregulation is reported, including breast cancer, esophageal squamous cell carcinoma, hepatocellular carcinoma, and oxidative stress-linked neurodegeneration. The near-haploid background ensures that observed phenotypes are primarily attributable to single-gene disruption.
Typical applications include western blot analysis of GPX7, GRP78, and CHOP; ROS quantification with H2DCFDA or CellROX; and cell viability assays under menadione or tunicamycin treatment. NRF2 pathway activity can be assessed via RT-qPCR for GCLC and HMOX1, and apoptosis can be measured by Annexin V/PI flow cytometry or immunofluorescence for ER stress markers. This model is ideal for dissecting NRF2 dynamics, performing drug sensitivity screens, and investigating GPX7 deficiency in cancer cell survival. For further technical information, contact Ascent Research.
