The ATP1A3 Knockout HAP1 Polyclonal Cells product consists of a CRISPR/Cas9-edited polyclonal knockout population targeting the human ATP1A3 gene in the HAP1 cell line. This heterogeneous pool of gene-disrupted cells serves as a robust loss-of-function model for the alpha-3 subunit of the Na+/K+-ATPase, enabling studies of its physiological roles in a controlled genetic background.
HAP1 is a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia line, exhibiting a male karyotype and BCR-ABL1 fusion. Its haploid nature facilitates efficient gene knockout and phenotypic interpretation by minimizing genetic redundancy. Widely used in genetic screens, HAP1 provides a stable and manipulable platform for studying gene function, though its hematopoietic origin should be considered when extrapolating to neuronal contexts.
ATP1A3 encodes the catalytic alpha-3 subunit of Na+/K+-ATPase, which hydrolyzes ATP to export three Na+ and import two K+ per cycle, establishing transmembrane electrochemical gradients. These gradients are essential for neuronal action potentials, neurotransmitter reuptake by SLC6 transporters (SLC6A1, SLC6A3), and secondary transport via NCX and Na+/H+ exchanger. The pump is regulated by PKA, PKC, and ouabain, and interacts with beta subunits (ATP1B1/2) and FXYD modulators (FXYD1, FXYD2), as well as cytoskeletal anchors ankyrin and spectrin.
Disruption of ATP1A3 in the haploid HAP1 background creates a powerful tool for dissecting Na+/K+-ATPase function and its relation to channelopathies. Mutations in ATP1A3 cause AHC, RDP, cerebellar ataxia, and CAPOS syndrome. While HAP1 is not neuronal, it allows high-throughput analysis of conserved cellular mechanisms such as ion homeostasis, volume regulation, and drug sensitivity, bypassing neuronal network complexities. This model also supports comparative studies among Na+/K+-ATPase isoforms.
Typical assays include western blotting, RT-qPCR, immunofluorescence, SBFI sodium imaging, ATPase activity measurements, ouabain sensitivity tests, and ionic stress viability assays. Key research applications encompass functional characterization of ATP1A3, drug screening for ion pump modulators, disease modeling, and isoform-specific investigations. For additional technical information, please contact Ascent Research.
