This product comprises a polyclonal population of HT29 human colorectal adenocarcinoma cells in which the ITPK1 gene has been disrupted via CRISPR/Cas9-mediated genome editing. The resulting polyclonal knockout cells provide a heterogeneous pool of loss-of-function variants, enabling robust functional studies without clonal selection biases. This model is designed for investigating ITPK1-dependent signaling in an intestinal epithelial cancer context.
The HT29 host cell line was originally derived from a colorectal adenocarcinoma of a 44-year-old female patient and exhibits epithelial morphology. HT29 cells are widely used as a model system for intestinal epithelial biology and colorectal cancer research, offering well-characterized growth properties, drug response profiles, and signaling pathway activities. Their adherent growth and ability to form polarized monolayers make them particularly suitable for studying inositol phosphate metabolism and calcium-mediated processes in epithelial cells.
ITPK1 encodes an inositol-1,3,4-trisphosphate 5/6-kinase that phosphorylates Ins(1,3,4)P3 to produce Ins(1,3,4,5)P4 and Ins(1,3,4,6)P4, thereby regulating inositol phosphate homeostasis and intracellular calcium dynamics. ITPK1 functions downstream of GPCR agonists and receptor tyrosine kinase ligands, and interacts with calmodulin and inositol polyphosphate multikinase (IPMK). Its activity modulates p53 protein stability, Akt phosphorylation, and ERK signaling, ultimately influencing apoptotic responses. The ITPK1 knockout disrupts this metabolic node, leading to altered levels of higher inositol phosphates such as IP5 and IP6, and perturbed p53-dependent apoptosis, which may impact colorectal cancer cell survival and chemoresistance.
In the context of HT29 colorectal cancer cells, ITPK1 knockout holds particular significance given the reliance of these cells on p53 status and inositol phosphate signaling for growth control and drug sensitivity. HT29 cells harbor wild-type p53, making them an ideal system for dissecting ITPK1??s role in p53 stabilization and apoptotic priming. Disruption of ITPK1 in this background can uncover mechanisms of chemoresistance, Ca2+ signaling dysregulation, and crosstalk between inositol phosphate metabolism and canonical cancer pathways such as Wnt/??-catenin signaling.
Researchers can employ this polyclonal knockout population to investigate inositol phosphate signaling in colorectal cancer using mass spectrometry-based profiling of inositol polyphosphates, calcium flux assays, and western blotting for p53, Akt, and ERK activation. The model is suitable for screening chemosensitizing agents, evaluating apoptosis via annexin V staining, and assessing colony formation under pharmacological challenge. Additional applications include CRISPR functional genomics, exploration of IP6-mediated pathways, and intestinal epithelial barrier studies. For further information or to discuss custom applications, please contact Ascent Research.
