The HMOX1 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population of HAP1 cells with disrupted HMOX1. This loss-of-function model enables investigation of heme oxygenase-1 in a near-haploid human background. The heterogeneous pool facilitates phenotypic assessments without clonal isolation.
HAP1 is a near-haploid fibroblast-like line derived from KBM-7 CML cells, retaining single copies of most chromosomes except disomy 8. Widely used for functional genomics, its simplified genetics support knockout studies by eliminating biallelic editing complexity, providing clear genotype-phenotype links.
HMOX1 catalyzes heme degradation to biliverdin, CO, and Fe2?, serving as a vital cytoprotective enzyme against oxidative stress. Expression is induced by Nrf2 (NFE2L2) via antioxidant response elements and repressed by Bach1. Products include biliverdin, rapidly reduced to antioxidant bilirubin by BLVRA, and CO, which signals through soluble guanylyl cyclase and inhibits caspase-3. Liberated iron upregulates ferritin (FTH1/FTL) for sequestration. HMOX1 interacts with cytochrome P450 reductase and p62/SQSTM1, integrating with Nrf2 activation loops.
In HAP1, HMOX1 disruption yields a hemizygous knockout, sensitizing cells to oxidative challenges and ferroptosis inducers. This model clarifies the gene’s role in Nrf2-mediated cytoprotection, iron handling, and CO signaling, leveraging the haploid background for unambiguous phenotypic readouts.
Applications include western blotting, RT-qPCR, and heme oxygenase activity assays. Cells can be stressed with hemin, H?O?, or erastin to assess ROS, viability, and ferroptosis. Co-IP with Nrf2, BLVRA, or p62 is feasible, and the model suits high-throughput screening for Nrf2/HMOX1 modulators. For further information, contact Ascent Research.
