The DPEP3 Knockout HAP1 Polyclonal Cells represent a diverse population of HAP1 human cells subjected to CRISPR/Cas9-mediated gene disruption targeting the DPEP3 locus. This genetically heterogeneous pool, generated without single-cell cloning, provides a robust loss-of-function model for investigating DPEP3-dependent processes. The polyclonal format captures a spectrum of editing outcomes across the cell population, enabling functional studies that reflect population-level gene ablation. As a research tool, these cells are suitable for applications where uniform knockout is not assumed, and they sidestep the biases inherent in clonal isolation. The product is delivered as a ready-to-use polyclonal knockout pool, facilitating immediate experimental deployment.
HAP1 is a near-haploid, fibroblast-like cell line originally derived from the male chronic myeloid leukemia line KBM-7. This adherent cell model features a predominantly haploid karyotype, with the exception of disomy for chromosome 8 and a portion of chromosome 15. The near-haploid state greatly enhances the efficiency of CRISPR-based knockout procedures, as disruption of a single allele is typically sufficient to eliminate gene function. HAP1 cells retain crucial cellular machinery, such as intact p53 signaling, and exhibit stable growth characteristics, making them a favored platform for genetic screens, drug target validation, and pathway dissection. Their leukemic origin also positions them as a relevant system for studying cancer-associated genes.
The DPEP3 gene encodes a glycosylphosphatidylinositol (GPI)-anchored membrane dipeptidase that belongs to the membrane dipeptidase family. The protein functions as a hydrolase that cleaves dipeptides, with demonstrated activity toward cysteinyl-bis-glycine, and catalyzes the conversion of the inflammatory lipid mediator leukotriene D4 to leukotriene E4. DPEP3 is embedded in lipid rafts via its GPI anchor, where it interacts with substrates and likely with raft-resident signaling molecules. By hydrolyzing leukotriene D4, DPEP3 modulates the balance of cysteinyl leukotrienes, which are potent bronchoconstrictors and pro-inflammatory agents. Additionally, the hydrolysis of cysteinyl-bis-glycine yields cysteine and glycine, thereby contributing to glutathione metabolism. Although upstream regulators of DPEP3 remain poorly defined, testis-enriched transcription factors such as NR5A1/SF-1 are thought to drive its expression in germ cells. The downstream products, leukotriene E4 and free amino acids, feed into downstream metabolic and signaling networks, linking DPEP3 to pathways of arachidonic acid metabolism, glutathione homeostasis, and peptide catabolism.
In the HAP1 context, disruption of DPEP3 offers a clean system to dissect its specific contributions to leukotriene signaling and dipeptide turnover. Given the near-haploid background, the polyclonal population retains editing heterogeneity while ensuring high penetrance of loss-of-function alleles across the pool. This makes the model particularly suited for studying DPEP3??s role in regulating intracellular glutathione levels, oxidative stress responses, and inflammatory mediator production. Moreover, because DPEP3 has been implicated in testicular cancer and leukemia, the HAP1 knockout platform allows exploration of its potential oncogenic or tumor-suppressive functions in a leukemia-derived cellular environment. Researchers can leverage this model to compare wild-type and knockout responses to chemotherapeutic agents or to oxidative challenges, shedding light on DPEP3??s involvement in drug resistance and cell survival.
These polyclonal knockout cells are ideal for a range of downstream assays, including Western blotting and RT-qPCR to confirm DPEP3 ablation, dipeptidase activity assays to measure enzymatic function, and LC-MS-based quantification of leukotriene and glutathione metabolites. Functional readouts such as cell viability and migration assays can be employed to assess the impact of DPEP3 loss on proliferation, survival, and motility. The model supports CRISPR knockout screening for synthetic lethal interactions, drug target validation, and metabolism-focused research. It also serves as a tool for investigating peptide catabolic processes and their contribution to cancer progression and inflammation. For additional technical specifications, pricing, and availability, please contact Ascent Research.
