The DPP3 Knockout HAP1 Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HAP1 human near-haploid cell line, designed for loss-of-function studies of DPP3. This polyclonal cell pool carries heterogeneous CRISPR-mediated disruptions in the DPP3 gene, providing a versatile tool for investigating DPP3-dependent biological processes without the clonal biases associated with single-cell-derived knockout lines. The polyclonal format ensures broad representation of knockout variants, enabling robust functional genomics analyses.
The HAP1 cell line is a near-haploid, adherent, fibroblast-like human cell line originally derived from the chronic myeloid leukemia KBM-7 line. Its haploid genome facilitates unambiguous genotype-phenotype correlations, making it an ideal host for CRISPR/Cas9-mediated knockout studies. HAP1 cells retain many characteristics of somatic cells, including responsiveness to oxidative and electrophilic stress, and are widely employed in high-throughput genetic screens and targeted gene disruption experiments. This genetic simplicity eliminates complications from diploid allele redundancy, ensuring that DPP3 gene disruption yields clear phenotypic outcomes.
Dipeptidyl peptidase 3 (DPP3) is a zinc-dependent aminopeptidase that cleaves dipeptides from oligopeptides, but its prominent role is as a positive regulator of the KEAP1-NRF2 antioxidant pathway. Under basal conditions, KEAP1 targets NRF2 for CUL3/RBX1-mediated ubiquitination and degradation. DPP3 directly binds KEAP1??s Kelch domain, competing with NRF2 and thereby stabilizing NRF2, which accumulates and activates transcription of cytoprotective genes (HMOX1, NQO1, GCLM) via ARE. Oxidative stress (H2O2) and electrophilic compounds like sulforaphane further upregulate DPP3, reinforcing this cytoprotective loop.
In the HAP1 haploid background, DPP3 disruption provides a clean loss-of-function model to dissect NRF2 signaling. HAP1 cells express functional KEAP1 and NRF2; thus, DPP3 ablation sensitizes cells to oxidative insults by impairing NRF2 stabilization, reducing downstream antioxidant defenses. This allows precise evaluation of DPP3??s role in redox balance and NRF2-dependent transcription, free from diploid allele interference. The polyclonal knockout population is a physiologically relevant platform for mechanistic studies.
The DPP3 Knockout HAP1 Polyclonal Cells are suited for functional studies of DPP3 in oxidative stress and cancer biology. Typical experiments involve H2O2 challenge combined with viability assays, ROS detection (DCFDA), and RT-qPCR analysis of NRF2 target genes (HMOX1, NQO1). The cells enable inhibitor screening using dipeptidyl peptidase activity measurement and NRF2 luciferase reporter assays, and facilitate peptide metabolism analysis. Western blotting for DPP3, KEAP1, and NRF2 confirms knockout efficiency and downstream pathway effects. For more information, please contact Ascent Research.
