The ATOX1 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal cell population derived from the human chronic myeloid leukemia (CML) cell line HAP1, engineered for loss-of-function studies of the ATOX1 gene. This product supplies a heterogeneous pool of knockout cells, enabling robust and reproducible assessment of ATOX1-dependent processes without the bias introduced by clonal isolation. As a polyclonal population, it is ideally suited for applications where bulk knockout phenotypes are desired, such as pooled genetic screens, biochemical assays, and high-throughput analyses. The cells are provided as a ready-to-use reagent, allowing immediate integration into copper biology and cancer research workflows.
HAP1 cells are a near-haploid human cell line originally derived from the KBM-7 CML line. They exhibit adherent growth, express the BCR-ABL oncogene, and retain a stable karyotype with a haploid chromosome complement for most chromosomes. This near-haploidy is a key advantage for CRISPR-based gene disruption, as a single targeting event can lead to complete loss of gene function, eliminating the need for biallelic editing required in diploid cells. HAP1 cells have become a cornerstone in functional genomics, extensively used in arrayed and pooled CRISPR screens, protein interaction mapping, and drug target validation. Their myeloid origin further positions them as a relevant model for leukemia-specific signaling and metal ion homeostasis studies.
ATOX1 encodes a metallochaperone that selectively binds cytosolic copper and shuttles it to the copper-transporting P-type ATPases ATP7A and ATP7B located at the trans-Golgi network. This copper delivery is essential for the activation of secreted cuproenzymes such as ceruloplasmin and for copper export from the cell. The chaperone function of ATOX1 is tightly regulated by intracellular copper availability and by oxidative stress, with the Nrf2 transcription factor serving as an upstream activator under redox challenge. Downstream, ATOX1-mediated copper transfer governs ATP7A and ATP7B trafficking and activity, thereby controlling systemic copper homeostasis. In addition to its biosynthetic role, ATOX1 contributes to antioxidant defense by sequestering copper and limiting Fenton-type generation of reactive oxygen species (ROS). The ATOX1-ATP7A/ATP7B axis thus represents a critical node linking metal ion regulation, redox balance, and cellular secretion.
In the HAP1 CML background, perturbation of ATOX1 creates a powerful model to dissect copper-dependent signaling and oxidative stress responses within a leukemic context. Copper metabolism is increasingly implicated in cancer proliferation and drug resistance, and chronic myeloid leukemia cells are known to exhibit altered redox states and metal ion dependencies. Disabling ATOX1 is predicted to impair copper delivery to ATP7A/ATP7B, leading to defects in copper excretion, loss of cuproenzyme activity, and increased vulnerability to copper-induced oxidative damage. This near-haploid knockout model therefore offers a clean genetic background to study how copper chaperone dysfunction affects leukemogenesis, cellular antioxidant capacity, and potential synthetic lethal interactions with pathways that further stress redox homeostasis.
The ATOX1 Knockout HAP1 Polyclonal Cells are a versatile tool for a wide array of functional investigations. Researchers can verify target gene disruption by Western blotting and RT-qPCR, and assess compensatory changes in copper transporters. Copper uptake and efflux assays, coupled with cell viability measurements under varying copper loads, provide quantitative readouts of copper handling defects. Immunofluorescence for ATP7A permits visualization of copper-dependent protein trafficking, while RNA-seq analyses reveal global transcriptomic adaptations to ATOX1 loss. The polyclonal nature of the population supports unbiased genetic interaction screens and drug modifier studies in a CML setting. For additional product details, pricing, or custom requests, please contact Ascent Research.
