The DPP9 Knockout Huh-7 Polyclonal Cells comprise a polyclonal population of CRISPR/Cas9-edited Huh-7 cells with targeted disruption of the DPP9 gene. This loss-of-function model enables stable abrogation of DPP9 serine protease activity without reliance on transient silencing or inhibitors, providing a robust tool for studying its cellular functions in a hepatocarcinoma background.
Huh-7 is a well-differentiated human hepatocellular carcinoma cell line originally isolated from a liver tumor in a 57-year-old Japanese male. It retains hepatocyte-like morphology and key metabolic enzymes, making it widely used for studies of liver cancer biology, hepatitis C virus replication, and drug metabolism. The line carries mutations in TP53 and CTNNB1, reflecting common oncogenic alterations in hepatocellular carcinoma.
DPP9 is an intracellular serine protease that cleaves N-terminal dipeptides from substrates with proline or alanine at the second position. It directly interacts with NLRP1 and the related protease DPP8 to negatively regulate the NLRP1 inflammasome, a multiprotein complex that activates caspase-1 and promotes IL-1?? secretion. DPP9 expression is upregulated by pro-inflammatory cytokines TNF-?? and IL-6 through NF-??B and STAT3 transcription factors. Downstream, DPP9 modulates levels of the chemokine CXCL10, inflammasome component NLRP1, and apoptosis regulators of the BCL2 family, linking immune signaling to cell death and metabolic pathways.
In the Huh-7 hepatic cancer context, DPP9 knockout disrupts inflammasome control and may alter apoptosis susceptibility, cytokine production, and metabolic homeostasis. This model allows investigation of how DPP9 influences tumor cell behavior and interactions with the immune microenvironment, particularly given the liver’s role in systemic metabolism and immune tolerance. The polyclonal nature maintains heterogeneity while ensuring robust knockout, suitable for functional assays.
Researchers can employ these cells for NLRP1 inflammasome studies using caspase-1 activity assays and IL-1?? ELISA, co-immunoprecipitation for DPP9-NLRP1 complexes, and flow cytometry to measure apoptosis. They also support RT-qPCR and Western blotting for downstream targets like CXCL10 and BCL2 members. Applications span cancer, inflammation, and metabolic disease. For further details, contact Ascent Research.
