The AOC1 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the AOC1 gene, which encodes the copper-containing amine oxidase, diamine oxidase. This heterogeneous pool of HAP1 cells harbors diverse mutations in AOC1, generated following Cas9-mediated double-strand breaks, providing a robust model for studying diamine oxidase deficiency in a near-haploid context without single-cell cloning.
The HAP1 cell line is a near-haploid, adherent human line derived from KBM-7 chronic myeloid leukemia, carrying a BCR-ABL fusion. With a haploid chromosome number (~25), HAP1 cells have single copies of most genes, facilitating CRISPR knockout studies and functional genomics and enabling clear genotype-phenotype links.
AOC1 encodes a homodimeric copper-dependent amine oxidase that catalyzes the oxidative deamination of histamine and putrescine, producing aldehydes, ammonia, and hydrogen peroxide. It is transcriptionally upregulated by inflammatory cytokines (TNF-??, IFN-??) and modulated by progesterone and allergen exposure. The enzyme reduces histamine receptor (HRH1, HRH2) signaling by degrading extracellular histamine and influences polyamine metabolism via putrescine, intersecting with ornithine decarboxylase (ODC1) and spermidine/spermine N1-acetyltransferase (SAT1). AOC1 interacts with copper chaperone ATOX1 and is functionally linked to catalase for peroxide detoxification. Loss of AOC1 leads to elevated histamine receptor activation, altered polyamine pools, oxidative stress, and potential cell cycle disruption.
In HAP1 cells, AOC1 knockout leverages the near-haploid genome for unambiguous dissection of diamine metabolism. The simplified genetic background makes this model particularly valuable for high-throughput screens to identify modulators of histamine sensitivity, polyamine homeostasis, or oxidative stress responses. The polyclonal population can be challenged with diamines to assess proliferation, ROS production, or aldehyde-mediated protein modifications. Additionally, the BCR-ABL-positive background provides a cancer-relevant context for exploring amine oxidase roles in oncogenic signaling.
Researchers can use these cells for studying histamine intolerance, allergic inflammation, polyamine metabolism in leukemia, screening diamine oxidase inhibitors, and gut barrier function. Compatible with diamine oxidase activity assays, histamine ELISA, polyamine HPLC, Western blot, RT-qPCR, proliferation (MTT/BrdU), and ROS (DCFDA) assays, this model supports comprehensive analyses. Knockout validation can be done by Sanger sequencing and immunoblotting. For support, contact Ascent Research.
