The APOM Knockout HAP1 Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the apolipoprotein M (APOM) gene in the HAP1 human cell line. This polyclonal pool provides a heterogeneous loss-of-function model, enabling robust functional investigations of APOM-dependent processes without clonal isolation.
HAP1 is a near-haploid human cell line originally derived from a male chronic myeloid leukemia patient. Its near-haploid karyotype, featuring a single copy of most chromosomes except a disomic region of chromosome 15, simplifies gene targeting and allows efficient generation of knockout models. HAP1 cells exhibit an adherent, fibroblast-like morphology and are widely employed in functional genomics and high-throughput genetic screens due to their stable culture characteristics.
APOM is a lipocalin family member predominantly associated with high-density lipoprotein (HDL) particles, where it acts as a specific chaperone for the bioactive sphingolipid sphingosine-1-phosphate (S1P). APOM expression is transcriptionally regulated by hepatocyte nuclear factor-1?? (HNF-1??), forkhead box protein A2 (FOXA2), and liver X receptor (LXR), with modulation by insulin and glucose. The APOM-S1P complex binds to S1P receptors (S1PR1-5) on target cells, initiating signaling cascades including AKT1-dependent phosphorylation of endothelial nitric oxide synthase (eNOS) to promote vasoprotection. APOM also interacts with HDL apolipoproteins (ApoA-I, ApoA-II), the endocytic receptor megalin, and scavenger receptor class B type I (SR-BI), linking HDL metabolism to anti-inflammatory and reverse cholesterol transport pathways.
In the HAP1 background, disruption of APOM eliminates endogenous HDL-associated S1P chaperoning, offering a simplified genetic platform to dissect HDL-S1P signaling without confounding hepatic or erythroid factors. The near-haploid genome ensures effective gene silencing, making this polyclonal model particularly valuable for studying lipid transport, endothelial barrier regulation, and the molecular underpinnings of atherosclerosis, coronary artery disease, type 2 diabetes, and sepsis.
Research applications encompass analysis of HDL function, S1P transport, and endothelial signaling through assays such as cholesterol efflux measurements, transwell endothelial permeability tests, S1P quantification by LC-MS/MS, co-immunoprecipitation of APOM with HDL, flow cytometric assessment of S1PR1 surface expression, and phospho-AKT/phospho-eNOS western blotting. For detailed protocols, dataset inquiries, or customization options, please contact Ascent Research.
