The ATP8A1 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HAP1 human haploid cell line. This product comprises a heterogeneous pool of edited cells in which the ATP8A1 gene has been disrupted via CRISPR/Cas9-mediated gene targeting, generating a loss-of-function model suitable for functional genomics and cell biology research. The polyclonal format avoids clonal artifacts and provides a representative population-level knockout phenotype, enabling robust and reproducible experimental designs.
The host HAP1 cell line is a near-haploid human cell model originally derived from the KBM-7 chronic myeloid leukemia cell line. Its haploid karyotype simplifies gene-editing efforts and facilitates the generation of null alleles through single guide RNA targeting. HAP1 cells are widely used in CRISPR-based functional screens, drug-target validation, and studies of fundamental cellular processes because the reduced genome complexity minimizes dominant-negative effects and enhances phenotype penetrance upon gene disruption.
ATP8A1 encodes an aminophospholipid flippase that actively translocates phosphatidylserine and phosphatidylethanolamine from the exoplasmic to the cytoplasmic leaflet of the plasma membrane, enforcing lipid asymmetry. This transport requires the cofactor CDC50A (TMEM30A) and is modulated by protein kinase C, phosphatidylinositol 4-phosphate, and calcium ions. ATP8A1 maintains inner-leaflet phosphatidylserine pools essential for KRas membrane localization, supports Akt and Rac1 signaling, and prevents aberrant phosphatidylserine exposure that triggers apoptosis. The flippase also functionally interacts with CDC50B and actin-binding proteins, linking lipid trafficking to cell polarity and survival.
Disruption of ATP8A1 in the HAP1 background provides a powerful system to dissect the molecular consequences of defective aminophospholipid translocation in a genetically tractable context. Loss of flippase activity leads to loss of membrane asymmetry, impaired KRas membrane anchorage, perturbed Akt and ERK pathway activation, and increased susceptibility to apoptosis, phenotypes that are readily interrogated in the haploid environment. The polyclonal knockout cell pool preserves genetic heterogeneity while uniformly lacking ATP8A1 function, making it ideal for studying the acute effects of lipid asymmetry loss on cancer cell signaling, membrane dynamics, and cell death regulation.
These cells are well-suited for a range of research applications including functional genomics, lipid trafficking and membrane asymmetry studies, apoptosis research, CRISPR screening, and drug target validation. Representative assays for characterizing the knockout phenotype include Western blotting, RT-qPCR, Annexin V flow cytometry, immunofluorescence microscopy, lipidomics, cell migration assays, phospho-signaling analysis, and co-immunoprecipitation. For additional technical information or to request a quotation, please contact Ascent Research.
