The ARG2 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from the near-haploid HAP1 cell line, engineered for disruption of the ARG2 gene encoding mitochondrial arginase II. This polyclonal knockout pool provides a robust loss-of-function model to study ARG2?Cdependent metabolic and signaling processes without requiring single-cell cloning. The targeted gene disruption eliminates the conversion of L-arginine to L-ornithine and urea, a key metabolic node that governs nitric oxide production, polyamine synthesis, and cell proliferation.
HAP1 is a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia background. Its haploid genome simplifies CRISPR/Cas9-mediated gene disruption, often yielding complete functional knockout and enabling clean genetic screens. Retaining leukemic signaling features, HAP1 is widely used for functional genomics and pathway dissection. The ARG2 knockout in this context allows unambiguous attribution of phenotypes to arginase II loss, supporting mechanistic studies in cancer metabolism and immune evasion.
ARG2, a mitochondrial enzyme, hydrolyzes L-arginine to L-ornithine and urea, competing with NOS isoforms for substrate and thus reducing NO generation. The product L-ornithine is further metabolized by ODC1 into polyamines (putrescine, spermidine, spermine) and by P5CS to proline, fueling proliferation and collagen synthesis. ARG2 expression is upregulated by IL-4, IL-13, and TGF-?? through STAT6/C/EBP??, and by hypoxia via HIF-1??; cAMP and glucocorticoids also modulate its levels. This arginine metabolic branch intersects with mTORC1 signaling and supports immune suppression and tumor growth.
In the HAP1 leukemia model, ARG2 disruption lifts substrate competition with NOS, potentially raising NO while depleting ornithine-derived metabolites??alterations that can impair proliferation and sensitize cells to apoptosis. This knockout pool enables LC-MS polyamine profiling, Griess assays for NO, and functional readouts such as MTS proliferation, Annexin V apoptosis, and Transwell migration assays. The haploid background strengthens the connection between ARG2 loss and observed phenotypes, making the model ideally suited for cancer metabolism research and inhibitor validation.
Applications include functional characterization of arginine metabolism, immune suppression mechanism studies, cancer drug resistance research, and CRISPR?based metabolic screening. The cells are compatible with Western blotting, RT-qPCR, arginase activity assays, RNA-seq, and metabolomics. They serve as a platform to validate ARG2 inhibitors and dissect metabolic vulnerabilities in oncology and immunology. For further details, please contact Ascent Research.
