The DPP9 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the DPP9 gene in the human HT29 colorectal adenocarcinoma cell line. This genetically heterogeneous pool carries targeted disruptions within the DPP9 locus, enabling loss-of-function studies without clonal selection. As a polyclonal knockout model, it reduces clonal artifacts and is well-suited for experiments requiring a mixed genetic background that more closely reflects the complexity of tumor cell populations.
The parental HT29 cell line is derived from a primary colon adenocarcinoma of a 44-year-old female and is widely employed as an in vitro model of intestinal epithelium. Notably, HT29 cells harbor well-defined oncogenic mutations in APC, KRAS, TP53, and PIK3CA, and are characterized as microsatellite stable and tumorigenic in immunocompromised mice. These genetic features make HT29 a valuable system for investigating colorectal cancer biology, drug transport, and epithelial barrier function, particularly in the context of tumor suppressor and oncogenic signaling pathways.
DPP9 encodes an intracellular serine protease that cleaves N-terminal dipeptides from substrates and acts as a critical negative regulator of the NLRP1 inflammasome. DPP9 binds to and inhibits NLRP1, preventing its oligomerization and the recruitment of the adaptor ASC. Upon DPP9 knockout, NLRP1 assembles into an active inflammasome platform, leading to caspase-1 activation and subsequent processing of pro-IL-1?? and pro-IL-18 into their mature, secreted forms. Active caspase-1 also triggers pyroptosis, a lytic, pro-inflammatory cell death. This cascade is modulated by upstream cytokines IFN-?? and TNF-??, which prime the inflammasome, and by the homologous peptidase DPP8 that partially substitutes for DPP9.
In the HT29 colorectal cancer model, DPP9 disruption provides a unique opportunity to study inflammasome regulation in a defined oncogenic context with mutations in APC, KRAS, TP53, and PIK3CA. This background allows exploration of cross-talk between oncogenic signaling and innate immunity. Because NLRP1-driven pyroptosis can influence tumor progression and therapeutic responses, these polyclonal knockout cells are valuable for investigating how inflammasome activation affects cancer cell survival and the tumor microenvironment, and for examining DPP9-dependent peptide processing in cancer cells.
Researchers can utilize these cells for diverse experimental approaches: Western blotting confirms DPP9 loss and caspase-1 cleavage; ELISA and LDH release assays quantify mature IL-1??/IL-18 and pyroptosis; RT-qPCR assesses transcript levels; and immunofluorescence visualizes ASC specks. The polyclonal knockout is ideal for screening DPP9 inhibitors, distinguishing apoptotic from pyroptotic death, and studying inflammasome dynamics under cytokine stimulation. For further information or to inquire about custom options, please contact Ascent Research.
