The ITPKC Knockout HT29 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma cell line, designed for loss-of-function studies of the ITPKC gene. This product leverages CRISPR/Cas9-mediated gene disruption to ablate ITPKC expression, generating a heterogeneous pool of knockout cells suitable for pooled functional analyses. The polyclonal format avoids clonal artifacts and captures population-level responses, enabling robust interrogation of ITPKC-dependent signaling and phenotypic changes in an epithelial colorectal cancer background.
The parental HT29 cell line was originally isolated from a 44-year-old female patient with colorectal adenocarcinoma and exhibits epithelial morphology. As a widely employed model for intestinal epithelial cell biology, HT29 cells have been instrumental in cancer research, drug development, and mechanistic studies of colorectal tumorigenesis. Their well-characterized growth behavior, differentiation capacity, and responsiveness to extracellular stimuli make them a reliable platform for investigating genes implicated in epithelial homeostasis and colorectal malignancy.
ITPKC encodes inositol-trisphosphate 3-kinase C, which phosphorylates IP3 to IP4, reducing IP3 levels and attenuating endoplasmic reticulum calcium release. This negatively regulates calcium-dependent signaling and proliferation. Activated downstream of phospholipase C by EGF, TNF-??, and GPCR agonists, ITPKC modulates effectors such as calcineurin/NFAT, CaMKII, and PKC, and interacts with calmodulin and IP3 receptors. By diminishing IP3-mediated calcium mobilization, ITPKC restrains the calcineurin/NFAT module and CaMKII cascade, limiting transcription of pro-proliferative and immune-related genes and influencing apoptosis.
In HT29 colorectal adenocarcinoma cells, ITPKC knockout allows dissection of calcium signaling roles in malignant epithelial growth. Deregulated calcium and NFAT activation are linked to colorectal cancer, and ITPKC loss may enhance calcium flux and NFAT-driven transcription, affecting proliferation, apoptosis, and migration. This model enables exploration of augmented calcium signaling in tumor phenotypes and testing of therapeutic targeting of inositol phosphate metabolism. It also facilitates investigation of ITPKC in colitis-associated cancer where calcium-dependent inflammatory signaling contributes.
The polyclonal knockout cells support diverse assays: Fluo-4 AM for calcium flux, NFAT luciferase reporters, MTT proliferation, Annexin V apoptosis, Transwell migration, and molecular profiling via Western blotting, RT-qPCR, and RNA-seq. Applications include studying calcium signaling in colorectal cancer, functional analysis of ITPKC in epithelial growth and apoptosis, drug screening targeting inositol phosphate metabolism, and exploring colitis-associated cancer mechanisms. For further information or specific experimental needs, please contact Ascent Research.
