APOA1 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from HAP1 human near-haploid leukemia cells. This product offers a loss-of-function model for the APOA1 gene, which encodes apolipoprotein A-I, the major structural protein of HDL. Generated through CRISPR/Cas9-mediated gene disruption at the APOA1 locus, the polyclonal format provides a diverse pool of cells with targeted mutations, suitable for pooled functional assays without clonal selection artifacts.
HAP1 cells originate from the KBM-7 chronic myeloid leukemia line and maintain a near-haploid karyotype, making them an optimal platform for gene-editing studies. The haploid genetic architecture facilitates clear genotype-phenotype relationships upon single-allele disruption. These cells are widely adopted for functional genomics, drug target screening, and signaling pathway analysis due to their robust proliferation and compatibility with high-throughput workflows.
APOA1 is a critical mediator of reverse cholesterol transport (RCT). It interacts with ABCA1 to accept cellular cholesterol and phospholipids, forming nascent HDL particles. As a cofactor for LCAT, APOA1 promotes cholesterol esterification, enabling HDL maturation. Mature HDL is recognized by SR-B1 on hepatocytes for selective cholesterol uptake. Transcriptional control of APOA1 involves HNF4A, PPARA agonists, LXRA/LXRB, and nuclear receptors such as ROR??. Downstream, APOA1-mediated HDL remodeling influences endothelial nitric oxide synthase activity and foam cell regression, underscoring its atheroprotective roles.
While APOA1 is not endogenously expressed at high levels in leukemic cells, the HAP1 knockout background is valuable for reconstitution experiments and interaction studies. When complemented with exogenous APOA1 or used in co-cultures with lipid-loaded cells, the model enables precise dissection of APOA1-dependent functions. The haploid setting ensures unambiguous interpretation of phenotypes, allowing assessment of APOA1??s interactions with ABCA1, LCAT, and SR-B1 in cholesterol efflux and HDL maturation.
Typical applications include cholesterol efflux assays, LCAT activity measurements, and SR-B1-mediated uptake studies. The polyclonal cells are amenable to FPLC-based HDL analysis, immunoblotting, RT-qPCR for APOA1 mRNA quantification, and lipidomics of sterol metabolism. In drug screening, they serve as a negative control for lipid-lowering agents and a platform to explore PPAR signaling modulators. Co-immunoprecipitation with ABCA1 or LCAT and endothelial function co-culture assays are further uses. For technical assistance, please contact Ascent Research.
