The ATP7B Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population originating from the HAP1 line, engineered to disrupt ATP7B gene function. This polyclonal knockout pool provides a genetically diverse loss-of-function model, avoiding clonal selection artifacts and enabling robust investigation of ATP7B-dependent copper transport mechanisms.
HAP1 is a near-haploid fibroblast-like cell line derived from chronic myeloid leukemia, serving as a hematopoietic progenitor cell model. Its near-haploidy simplifies gene targeting and phenotypic analysis, as each gene is typically present in a single copy. The cells exhibit adherent growth and stable culture properties, making them ideal for high-throughput functional genomics and reproducible experimentation.
ATP7B encodes a copper-transporting P-type ATPase that mediates copper excretion into bile and incorporation into ceruloplasmin. Transcriptionally, ATP7B is induced by copper binding to the transcription factor MTF1. On the protein level, ATP7B receives copper from the chaperone ATOX1 and is regulated by COMMD1. Within the cellular copper network, ATP7B functions in concert with ATP7A, additional chaperones (CCS, COX17), ceruloplasmin, and metallothioneins. Its disruption abolishes copper efflux, prevents ceruloplasmin maturation, and leads to intracellular copper accumulation and associated oxidative stress.
In the HAP1 near-haploid background, deletion of the single ATP7B allele creates a clean knockout model that recapitulates key features of Wilson disease. This system allows precise analysis of copper-induced signaling cascades, interrogation of interactions with ATOX1 and COMMD1, and investigation of potential compensatory responses by ATP7A and metallothioneins. The simplified genome eliminates confounding heterozygous effects, enhancing the clarity of copper toxicity studies.
Researchers can utilize these polyclonal knockout cells for Wilson disease pathogenesis studies, copper metabolism research, and high-throughput drug screening for copper overload disorders. Feasible assays include copper content measurement by atomic absorption spectroscopy, ceruloplasmin enzymatic activity assays, copper efflux kinetics, and cell viability under copper stress. Gene disruption verification can be performed by RT-qPCR and immunofluorescence. These cells also provide a suitable platform for gene therapy vector testing and CRISPR-mediated rescue experiments. For additional product information, please contact Ascent Research.
