The DPEP2 Knockout HAP1 Polyclonal Cells product consists of a polyclonal population of HAP1 cells that have undergone CRISPR/Cas9-mediated disruption of the DPEP2 gene, which encodes a membrane-bound dipeptidase. This polyclonal knockout cell population provides a physiologically relevant loss-of-function model for investigating DPEP2-dependent dipeptide hydrolysis and its role in leukotriene metabolism. By targeting DPEP2 in a near-haploid background, the cells offer a simplified genetic context for studying gene function without the confounding effects of a second allele, making them ideal for functional genomics and pathway dissection. The polyclonal nature ensures that the population retains genetic diversity, which can be advantageous for capturing a range of knockout phenotypes and for downstream applications that benefit from heterogeneous cellular responses.
HAP1 is a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia line, originally isolated from a 40-year-old male patient. Its haploid karyotype renders it particularly useful for genetic perturbation studies, as recessive mutations are immediately phenotypically apparent, enabling straightforward gene-trait associations. HAP1 cells are widely employed as a model system in drug discovery, target validation, and functional genomics due to their ease of genetic manipulation, rapid growth, and compatibility with a broad array of cell-based assays. This cell line provides a robust platform for generating knockout models that yield clear and interpretable results, especially when studying genes involved in complex signaling cascades like those governing inflammation.
DPEP2 functions as a membrane dipeptidase that specifically hydrolyzes dipeptides, including the cysteinyl leukotriene leukotriene D4 (LTD4). In the leukotriene signaling cascade, LTD4 is a potent pro-inflammatory mediator that acts through cysteinyl leukotriene receptors CysLTR1 and CysLTR2 to trigger intracellular calcium mobilization and downstream inflammatory responses. DPEP2 converts LTD4 to the less active leukotriene E4 (LTE4), thereby terminating signaling. The expression of DPEP2 is regulated by cytokines such as IL-4 and IL-13, as well as the NF-??B pathway. By interacting with glutathione and other dipeptidases, DPEP2 is positioned at a critical junction within arachidonic acid metabolism and glutathione-linked pathways. Disruption of DPEP2 leads to accumulation of LTD4 and sustained cysteinyl leukotriene receptor activation, resulting in enhanced and prolonged inflammatory signals.
Eliminating DPEP2 dipeptidase activity in HAP1 cells impairs the degradation of LTD4, mimicking conditions of heightened cysteinyl leukotriene signaling observed in chronic inflammatory disorders such as asthma, allergic inflammation, and chronic obstructive pulmonary disease. This polyclonal knockout model enables researchers to dissect the specific contributions of DPEP2 to leukotriene metabolism without the genetic redundancy present in diploid cells. Because HAP1 cells are derived from a hematopoietic lineage, they also offer a contextually relevant host for studying inflammation-related processes, including chemotaxis and cytokine responses. The model can be used to evaluate how loss of DPEP2 modulates downstream targets like LTE4 production and calcium flux, as well as to screen for modulators that restore normal leukotriene turnover.
Typical applications of these DPEP2 knockout polyclonal cells include functional genomics screens to identify novel regulators of dipeptidase activity, target validation studies for anti-inflammatory drug development, and mechanistic investigations into leukotriene signaling pathways. They can be employed in a variety of quantitative assays, such as Western blotting to verify loss of DPEP2 protein, RT-qPCR to confirm transcript ablation, LTD4 hydrolysis assays to measure enzymatic function, calcium flux assays to assess receptor-mediated signaling, chemotaxis assays to evaluate cell migration, flow cytometry to profile inflammatory marker expression, and RNA-seq for transcriptome-wide analysis of altered gene expression. These cells serve as a versatile platform for both basic research and preclinical therapeutic testing. For further information or customized applications, please contact Ascent Research.
