DPEP1 Knockout HAP1 Polyclonal Cells are a pool of HAP1 cells that have undergone CRISPR/Cas9-mediated disruption of the DPEP1 gene. This polyclonal population contains diverse DPEP1 loss-of-function alleles, providing a genetically heterogeneous knockout model suitable for pooled screening and bulk functional assays.
HAP1 is a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia cell line. Originating from a male donor, HAP1 cells retain a stable haploid karyotype that facilitates efficient gene knockout and functional genomic studies. Their rapid growth and ease of genetic manipulation make HAP1 a widely adopted model for cancer research, drug target validation, and large-scale genetic screens.
DPEP1 encodes a glycosylphosphatidylinositol (GPI)-anchored dipeptidase that hydrolyzes dipeptides and converts the pro-inflammatory lipid mediator leukotriene D4 (LTD4) to leukotriene E4 (LTE4). This enzymatic activity is part of the arachidonic acid metabolism and glutathione metabolic pathways, where DPEP1 acts downstream of 5-lipoxygenase and LTC4 synthase. DPEP1 is transcriptionally regulated by the SP1 transcription factor and is responsive to glucocorticoids and inflammatory cytokines. Its interaction with substrates such as LTD4 and cystinyl-bis-glycine, along with extracellular matrix components, influences leukotriene signaling through CysLT receptors (G protein-coupled receptors) and modulates extracellular matrix remodeling and cell adhesion. Additionally, DPEP1-mediated dipeptide hydrolysis releases cysteine and contributes to glutathione homeostasis, linking its function to oxidative stress responses.
In the context of the HAP1 near-haploid background, DPEP1 knockout provides a clean loss-of-function model that allows unambiguous dissection of DPEP1-dependent phenotypes. This is particularly valuable for investigating the role of leukotriene metabolism in cancer cell motility and invasion, as DPEP1 has been implicated in promoting metastasis in colorectal cancer and renal cell carcinoma. The polyclonal nature of the knockout population enables pooled functional screens, such as CRISPR drop-out assays, and provides a robust system for evaluating drug responses without clonal bias.
Typical research applications include quantitative analysis of leukotriene metabolites via LC-MS, Transwell migration and invasion assays to assess metastatic potential, and cell adhesion assays to examine interactions with extracellular matrix proteins. The DPEP1 knockout HAP1 polyclonal cells are also suitable for high-throughput drug screening targeting the leukotriene pathway and for functional genomics studies using haploid genetics. Western blotting and RT-qPCR can be employed to confirm DPEP1 disruption and to monitor downstream effects on gene expression. For further information and to explore custom applications, please contact Ascent Research.
