The DNPEP Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colon adenocarcinoma cell line, designed to disrupt the DNPEP gene encoding aspartyl aminopeptidase. This polyclonal population provides a powerful loss-of-function model for investigating DNPEP biology without requiring single-cell cloning, enabling researchers to assess pooled knockout effects across a population of edited cells.
The HT29 host cell line is a widely used model of human colorectal adenocarcinoma, originally isolated from a female donor. As adherent epithelial cells, HT29 cells recapitulate key aspects of intestinal epithelial physiology and are extensively employed in colorectal cancer research, including studies on cell signaling, metabolism, and drug response. Their robust growth and well-characterized molecular features make them an ideal background for gene-editing applications.
DNPEP encodes an aminopeptidase that specifically cleaves N-terminal aspartate and glutamate residues from peptide substrates, playing a critical role in protein catabolism, amino acid recycling, and peptide hormone inactivation. Mechanistically, DNPEP functions downstream of the renin-angiotensin system, converting angiotensin II to angiotensin III, and influences mTORC1 signaling through modulation of intracellular amino acid availability. DNPEP activity is regulated by nutrient deprivation, mTORC1 signaling, and the SP1 transcription factor, and it interacts with other aminopeptidases such as ANPEP and LAP3. Disruption of DNPEP therefore blocks the removal of N-terminal acidic amino acids, impairing peptide hormone processing and altering cellular metabolic pathways.
In the context of colorectal cancer, DNPEP knockout in HT29 cells provides a physiologically relevant platform to dissect the role of aspartyl aminopeptidase in tumor metabolism and signaling. Loss of DNPEP disrupts local angiotensin II degradation and amino acid recycling, potentially affecting mTORC1-dependent proliferation and stress responses. This model is particularly valuable for exploring links between aminopeptidase activity and pathologies such as hypertension, metabolic syndrome, and cancer cachexia, where dysregulated peptide hormone metabolism and amino acid homeostasis are implicated.
Researchers can utilize this knockout model for a broad range of functional studies, including assessment of aminopeptidase activity using Asp-AMC substrates, angiotensin II degradation assays, and quantification of intracellular amino acid pools. The polyclonal population is suitable for downstream applications such as proliferation and migration assays, drug sensitivity screens, and RNA-seq?Cbased transcriptomic profiling. By enabling detailed investigation of DNPEP-dependent processes in colon cancer cells, this product supports advances in understanding peptide hormone metabolism, amino acid sensing, and therapeutic targeting. For further information or to acquire these cells, please contact Ascent Research.
