The ATP11B Knockout HAP1 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal population designed for functional disruption of the ATP11B gene in the HAP1 host cell background. This polyclonal knockout model provides a robust loss-of-function system for investigating the role of ATP11B in phospholipid transport and membrane asymmetry. Generated through CRISPR/Cas9-mediated gene disruption, the product eliminates the need for single-cell cloning and permits direct analysis of a heterogeneous knockout pool, making it well-suited for phenotypic screening and pathway dissection.
The host cell line HAP1 is a near-haploid, adherent human cell line derived from the chronic myeloid leukemia cell line KBM-7. It retains the BCR-ABL translocation characteristic of CML and presents a male karyotype with disomy for chromosome 8 and a portion of chromosome 15. The near-haploid state minimizes genetic complexity and facilitates gene targeting and functional genomics studies, rendering HAP1 an ideal platform for generating clean knockout models and interpreting genotype?Cphenotype relationships with reduced confounding from heterozygous alleles.
ATP11B encodes a P4-type ATPase phospholipid flippase that, in complex with its obligate beta subunit CDC50A (TMEM30A), actively translocates aminophospholipids??phosphatidylserine (PS) and phosphatidylethanolamine (PE)??from the exoplasmic to the cytoplasmic leaflet of the plasma membrane. This activity maintains transbilayer lipid asymmetry, prevents aberrant exposure of PS on the cell surface, and supports membrane curvature regulation. The flippase is subject to regulatory cleavage by caspases (notably caspase-3) during apoptosis and is responsive to calcium signaling, linking ATP11B activity directly to apoptotic progression and cellular signaling cascades that depend on membrane lipid distribution.
In the HAP1 context, ATP11B knockout disrupts inward phospholipid translocation, leading to spontaneous and sustained externalization of PS. Because HAP1 cells are near-haploid, a single CRISPR-targeted allele is sufficient to abolish flippase function, yielding a clear and immediate phenotype. This PS exposure can be quantified by high-sensitivity assays such as Annexin V binding and flow cytometry, and the defect has implications for apoptotic cell clearance, membrane trafficking, and drug resistance mechanisms. The model thus allows researchers to dissect how loss of lipid asymmetry influences cell fate decisions and therapeutic responses.
This ATP11B knockout polyclonal population is suitable for a wide range of applications including apoptosis signaling analysis, membrane biology investigations, drug resistance profiling, and high-content phenotypic screens. Commonly employed assays with this model include Annexin V staining and flow cytometric measurement of surface PS, Western blotting for ATP11B and cleaved caspase-3, RT-qPCR to verify transcript disruption, lipidomics to assess global phospholipid composition, and cell viability or apoptosis assays. For further information on this product or to explore custom applications, please contact Ascent Research.
