The ATP1B1 Knockout HAP1 Polyclonal Cells product consists of a CRISPR/Cas9-edited polyclonal population carrying targeted disruptions in the ATP1B1 gene, generating a heterogeneous loss-of-function model. This polyclonal format retains the natural diversity of knockout alleles produced by non?homologous end joining, minimizing clonal artifacts and enhancing the robustness of functional studies. Researchers can utilize this population to investigate the multifaceted roles of ATP1B1 without the constraints of single?clone bias.
HAP1 is a human near?haploid fibroblastoid cell line derived from the KBM?7 chronic myeloid leukemia isolate. Its near?haploid karyotype simplifies genetic analyses and has established it as a preferred model for high?throughput genetic screens and drug target discovery. The cells grow adherently and maintain a stable male genetic background, supporting reproducible experiments in cancer biology and signal transduction.
ATP1B1 encodes the beta?1 subunit of the Na?/K??ATPase, an integral plasma membrane pump that forms an obligatory heterodimer with the alpha subunit (ATP1A1) to actively transport sodium out of and potassium into cells. This ion translocation maintains electrochemical gradients, membrane potential, and secondary transport. ATP1B1 also scaffolds a signaling microdomain by interacting with Caveolin?1, Src kinase, Ankyrin, and FXYD proteins. Transcriptional regulation involves SP1, aldosterone, T3, and cAMP/PKA. Consequently, ATP1B1 controls the activities of the sodium?calcium exchanger NCX1 and Src kinase, which in turn engages MAPK1/3 to influence proliferation and survival. Knockout of ATP1B1 disrupts ion homeostasis, depolarizes the membrane, and impairs Src?MAPK signaling.
In the HAP1 leukemic background, deletion of ATP1B1 reveals how Na?/K??ATPase?dependent signaling modulates cancer cell behavior. The loss disrupts intracellular Na? and K? balance, depolarizes the membrane potential, and attenuates Src?mediated signaling, thereby affecting adhesion and mitogenic responses. This model is especially valuable for probing the intersection of ion transport dysfunction and oncogenic pathways and for evaluating the selectivity of cardiac glycosides such as ouabain that target the Na?/K??ATPase.
This polyclonal knockout population supports a broad panel of functional assays. Western blotting and RT?qPCR verify ATP1B1 ablation, while ouabain sensitivity assays, SBFI?based intracellular Na? measurements, and DiBAC4(3) membrane potential recordings characterize ionic phenotypes. Co?immunoprecipitation with ATP1A1 and phospho?Src analysis probe signaling complex integrity, and MTT assays assess cell viability. Applications range from studying ion transport disorders and cancer cell signaling to cardiac electrophysiology and drug target validation for cardiac glycosides. For further technical information or to place an order, please contact Ascent Research.
