The ATP11C Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal population in which the ATP11C gene has been disrupted. This knockout model is established in the HAP1 host background, providing a versatile system for studying the loss of ATP11C function. The polyclonal format ensures a broad spectrum of gene-editing events, facilitating robust phenotypic analyses without selection bias inherent to clonal isolates.
HAP1 is a near-haploid fibroblast-like cell line derived from chronic myeloid leukemia. Its near-haploid karyotype enables efficient gene targeting, as a single editing event can abolish gene function, making it an ideal platform for CRISPR/Cas9 knockout studies. The cell line retains key signaling pathways, including those governing apoptosis and membrane dynamics, supporting physiologically relevant investigations.
ATP11C encodes a P4-ATPase phospholipid flippase that actively transports phosphatidylserine to the inner leaflet of the plasma membrane, maintaining membrane asymmetry. This activity is mediated by its heterodimeric partner CDC50A (TMEM30A) and is regulated by calcium ions (activator) and caspase-3 cleavage (inactivator). Loss of flippase function leads to phosphatidylserine externalization, a hallmark of apoptosis and efferocytosis, and disrupts B cell receptor signaling. ATP11C plays a critical role in lymphocyte development and iron homeostasis, and its deficiency is associated with X-linked congenital dyserythropoietic anemia and B-cell lymphopenia.
In HAP1 cells, ATP11C knockout abolishes flippase activity, resulting in constitutive phosphatidylserine exposure and impaired apoptosis signaling. This model recapitulates key cellular phenotypes of ATP11C-related disorders, such as defective erythropoiesis and B cell maturation. The disruption clarifies the signaling interplay between ATP11C, CDC50A, and caspases, and their downstream effects on pathways involving the B cell receptor and erythropoietin receptor. The near-haploid background ensures unambiguous genotype-phenotype correlations.
Researchers can utilize this polyclonal knockout population in diverse functional assays, including Annexin V staining for phosphatidylserine exposure, flow cytometry for apoptosis, and flippase activity measurements. Biochemical characterization via Western blot for ATP11C and CDC50A, along with immunofluorescence for membrane asymmetry, is readily achievable. The cells also support B cell differentiation assays and erythroid colony formation, making them suitable for modeling X-linked anemia and screening flippase-targeted compounds. For further information, contact Ascent Research.
