The DNPEP Knockout HCT 116 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal population with targeted disruption of the DNPEP gene, which encodes aspartyl aminopeptidase. This enzyme cleaves N-terminal aspartate and glutamate residues from peptide substrates, including neuropeptides and angiotensin. The polyclonal format provides a heterogeneous pool of edited cells, avoiding clonal selection bias and reflecting population-level genetic effects. This knockout model is designed for loss-of-function studies in a colorectal carcinoma background.
Derived from HCT 116 human colorectal carcinoma epithelial cells, the host line exhibits microsatellite instability-high (MSI-H) due to MLH1 promoter hypermethylation and harbors oncogenic mutations in KRAS (G13D) and CTNNB1 (??-catenin). These alterations constitutively activate MAPK/ERK and Wnt/??-catenin pathways, establishing a well-defined model for studying signal transduction in cancer. The combination of MSI-H and driver mutations makes this system particularly valuable for exploring genotype-phenotype relationships upon aminopeptidase ablation.
DNPEP functions as a cytosolic aminopeptidase that hydrolyzes N-terminal aspartyl and glutamyl residues from bioactive peptides, thereby regulating hormone and neuropeptide levels. Within the renin-angiotensin system, DNPEP processes angiotensin II downstream of renin and ACE, influencing AT1R-mediated signaling. Cross-talk with Wnt and MAPK pathways is suggested, as ??-catenin and KRAS/ERK are hyperactive in HCT 116 cells, and DNPEP-mediated peptide turnover may modulate these cascades. Loss of DNPEP likely alters the peptide substrate pool, impacting signal transduction through AT1R, MEK-ERK, and TCF/LEF transcriptional programs.
Knockout of DNPEP in the HCT 116 background creates an experimental system to evaluate how aminopeptidase activity integrates with oncogenic signaling. The pre-existing dysregulation of Wnt and MAPK enables assessment of whether DNPEP loss exacerbates or suppresses tumorigenic phenotypes, particularly in the context of peptide hormone metabolism. This model is relevant for investigating the role of angiotensin and other neuropeptides in tumor cell proliferation, migration, and microenvironmental interactions.
Applications include aminopeptidase activity assays, Western blotting for phospho-ERK and ??-catenin, RT-qPCR of Wnt target genes (e.g., AXIN2, MYC), ELISA-based angiotensin II quantification, and cell proliferation or migration studies. The polyclonal pool is well-suited for drug target validation, especially for agents targeting the renin-angiotensin system or aminopeptidase activity in colorectal cancer. For additional information or technical support, please contact Ascent Research.
