The ATP7A Knockout HAP1 Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal population with disrupted ATP7A gene in a near-haploid human background. This loss-of-function model allows study of ATP7A-dependent copper homeostasis without clonal artifacts, as the heterogeneous pool eliminates wild-type protein expression across the culture.
HAP1 is a male-derived, near-haploid cell line from KBM-7 chronic myeloid leukemia. Its single chromosome set facilitates functional genomics by enabling population-level null phenotypes upon gene disruption. Immortalized hematopoietic origin supports robust growth and is relevant for copper biology in a transformed cellular context.
ATP7A is a copper-transporting ATPase that loads secretory cuproenzymes (e.g., lysyl oxidase, tyrosinase) via copper delivery to the trans-Golgi network, utilizing the chaperone ATOX1. Under high copper, it shifts to the plasma membrane for efflux, regulated by COMMD1 and AP-2. Expression is controlled by copper, SP1, and hypoxia, and downstream effects include modulation of cellular copper levels and oxidative stress. It operates with the importer CTR1, the related exporter ATP7B, chaperone CCS, and ceruloplasmin.
The near-haploid HAP1 context ensures that ATP7A disruption results in uniform functional loss, ideal for reproducible copper sensitivity assays and redox measurements. This system is valuable for investigating copper??s role in hematopoietic cells and leukemic proliferation, with streamlined genetics enabling moderate-throughput compound screens.
Applications include Menkes disease modeling, cancer sensitivity to copper chelators, and oxidative stress analysis. Readouts can employ Western blot, RT-qPCR, immunofluorescence, ICP-MS for copper content, and flow cytometry for ROS. This versatile tool aids research into copper homeostasis, metal toxicity, and therapeutic targeting. Contact Ascent Research for details.
