The DPP7 Knockout HT29 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma cell line, engineered to disrupt the DPP7 gene. This pooled population retains heterogeneous genomic edits, offering a robust loss-of-function model for studying DPP7 without clonal selection bias. By abolishing DPP7 expression, these cells enable investigations into lysosomal dipeptidyl peptidase activity and its broader biological roles.
HT29 cells, isolated from a 44-year-old female with colorectal adenocarcinoma, serve as a well-established model of human intestinal epithelium. These epithelial cells maintain the capacity to differentiate into enterocyte-like and mucus-producing goblet cell phenotypes under appropriate culture conditions, making them particularly relevant for studies of intestinal physiology and colorectal cancer biology. Their use in biomedical research spans nutrient transport, barrier function, and tumor microenvironment interactions.
DPP7 encodes a lysosomal serine protease that selectively cleaves N-terminal dipeptides from peptide substrates, with a preference for penultimate proline or alanine residues. Its catalytic activity is regulated by upstream signals, including transcription factor EB (TFEB), TGF-?? signaling, and nutrient-sensitive pathways such as serum starvation. Within lysosomes, DPP7 cooperates with cathepsin B and cathepsin L to complete terminal protein degradation. Moreover, DPP7 contributes to MHC class II antigen processing by generating peptide fragments and modulating the cleavage of neuropeptides such as substance P, neuropeptide Y, and bradykinin, thereby influencing downstream immunomodulatory and inflammatory responses.
In the context of HT29 colorectal cancer cells, disruption of DPP7 is predicted to impair lysosomal proteolytic capacity and alter the repertoire of peptides available for MHC class II loading. This may compromise immune surveillance mechanisms and affect tumor cell homeostasis, apoptosis, and proliferation. Consequently, the knockout model provides a physiologically relevant platform to dissect how lysosomal dysfunction intersects with oncogenic signaling and antigen presentation in intestinal epithelial cells, offering insights into the interplay between protein catabolism and immune recognition.
These polyclonal knockout cells are suitable for a broad range of functional assays, including Western blotting and RT-qPCR to confirm DPP7 ablation, fluorogenic dipeptidyl peptidase activity measurements, and flow cytometric analysis of MHC class II surface expression. They enable proteomic profiling of accumulated substrates via mass spectrometry and allow functional antigen presentation assays using T cell readouts. Furthermore, the model supports high-content screening of DPP7 inhibitors and combinatorial studies with chemotherapeutics. Proliferation and apoptosis assays (MTT, Annexin V/PI) together with immunofluorescence staining for lysosomal markers such as LAMP-1 can be employed to assess the cellular consequences of DPP7 loss. For detailed technical specifications, please contact Ascent Research.
