The EGLN1 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the EGLN1 gene, which encodes prolyl hydroxylase domain-containing protein 2 (PHD2). This polyclonal pool comprises a mixed population of edited cells, providing a robust loss-of-function model for investigating the hypoxia-inducible factor (HIF) signaling pathway and oxygen sensing without the need for single-cell cloning.
These cells utilize the HAP1 cell line, a near-haploid human chronic myeloid leukemia (CML) model originally derived from KBM-7. HAP1 cells grow as an adherent monolayer, are p53-deficient, and retain a stable haploid karyotype, making them exceptionally suited for genetic screening and cancer biology applications. The haploid genome simplifies CRISPR/Cas9-mediated knockout generation, often yielding full functional gene loss in a single step.
EGLN1 functions as a primary oxygen sensor by catalyzing prolyl hydroxylation of HIF-1?? and HIF-2??. Under normoxia, this modification promotes interaction with the von Hippel-Lindau (VHL) E3 ubiquitin ligase complex, which includes Elongin B, Elongin C, Cul2, and Rbx1, leading to HIF-?? ubiquitination and proteasomal degradation. EGLN1 activity depends on oxygen, 2-oxoglutarate, iron, and ascorbate, and is inhibited by succinate, fumarate, and reactive oxygen species. During hypoxia, hydroxylation is suppressed, stabilizing HIF-??, which then translocates to the nucleus, dimerizes with ARNT, and induces target genes such as VEGFA and SLC2A1, regulating angiogenesis, erythropoiesis, and metabolism.
In the p53-deficient, haploid HAP1 background, EGLN1 knockout offers a clean system for dissecting HIF pathway dynamics without interference from p53-mediated stress responses. The polyclonal nature of the knockout ensures population-level loss of function, allowing researchers to examine HIF-1?? accumulation, transcriptional responses, and biochemical interactions under controlled oxygen conditions. This model is ideal for comparing normoxic and hypoxic states and for screening pharmacological inhibitors of prolyl hydroxylases.
Typical applications include hypoxia signaling research, HIF pathway analysis, cancer hypoxia studies, PHD inhibitor drug screening, erythropoiesis regulation, metabolic adaptation, and ischemic disease modeling. This cell product enables assays such as HIF-1?? western blotting, RT-qPCR of target genes (e.g., VEGFA, SLC2A1), luciferase reporter assays, co-immunoprecipitation, and functional tests like proliferation, migration, and colony formation under hypoxia. For further information or support, please contact Ascent Research.
