The ATP1B3 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population featuring disruption of the ATP1B3 gene. This product consists of a heterogeneous pool of HAP1 cells carrying diverse loss-of-function mutations, offering a robust model for functional genomics without clonal biases. The polyclonal format minimizes clone-specific artifacts and is well-suited for a range of downstream assays in ion transport and cancer research.
HAP1 is a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia line. Its haploid karyotype simplifies gene knockout, requiring modification of a single allele. HAP1 cells are widely used for genetic manipulation, drug target validation, and high-throughput screening thanks to their stable growth and tractability. The leukemic origin further provides a context for studying cancer-related signaling pathways.
ATP1B3 encodes the ??3 subunit of the Na+/K+-ATPase, a plasma membrane pump essential for maintaining Na+ and K+ gradients. The ??3 subunit interacts with ?? subunits (ATP1A1, ATP1A2, ATP1A3) and adaptor proteins such as caveolin-1, ankyrin, and FXYD proteins to form functional complexes. ATP1B3 is transcriptionally regulated by hormones like aldosterone and thyroid hormone and modulated by cAMP/PKA, protein kinase C, and insulin. It directly interacts with SRC kinase, and its disruption leads to aberrant activation of MAPK1/3 (ERK1/2) and EGFR signaling, linking ion homeostasis to cell proliferation and survival.
In the near-haploid HAP1 background, ATP1B3 disruption provides an unambiguous loss-of-function model. Haploidy ensures that the polyclonal population collectively abolishes protein function, enabling clear phenotype interpretation. This model is particularly powerful for dissecting how ATP1B3 loss impacts cell volume regulation, membrane potential, and SRC/ERK pathway activation. Given the leukemic context, it also facilitates investigation of ion gradient-driven malignant properties, including proliferation and drug resistance, and contributes to understanding diseases such as Charcot-Marie-Tooth disease, hypertension, and renal tubular acidosis.
Applications encompass western blotting for ATP1B3 and ATP1A1, Na+/K+-ATPase activity assays, intracellular ion concentration measurements, cell volume monitoring, and proliferation assays. Signaling output can be assessed via phospho-ERK ELISA and SRC kinase activity assays, while transcriptomic approaches like RNA-seq and RT-qPCR reveal pathway-level adaptations. This versatile knockout model supports target validation, ion physiology studies, and cancer signaling research. For further details, please contact Ascent Research.
