The ITPKA Knockout HCT 116 Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout cell population generated from the HCT 116 human colorectal carcinoma cell line, with disruption of the ITPKA gene to establish a loss-of-function model. This polyclonal format preserves genetic diversity, reducing the risk of clonal selection biases and more faithfully representing the heterogeneity typical of tumor cell populations. By eliminating ITPKA expression, these cells provide a versatile platform for dissecting the gene??s contributions to key cellular processes.
HCT 116 is a widely used colorectal carcinoma cell line with epithelial morphology, known to carry an activating KRAS G13D mutation and a loss-of-function TP53 mutation. These genetic lesions drive constitutive MAPK pathway activity, suppress p53-dependent checkpoints, and contribute to aggressive tumorigenic traits such as enhanced proliferation and resistance to apoptosis. Its well-documented genomics and reproducible in vitro growth characteristics make HCT 116 an established model for studying oncogenic signaling and anticancer drug discovery.
ITPKA encodes inositol-trisphosphate 3-kinase A, a Ca2+/calmodulin-dependent enzyme that phosphorylates inositol 1,4,5-trisphosphate (IP3) to generate inositol 1,3,4,5-tetrakisphosphate (IP4). This reaction terminates IP3-mediated calcium release from intracellular stores while producing IP4, a second messenger that regulates Akt signaling and actin cytoskeleton reorganization. ITPKA is activated downstream of epidermal growth factor receptor (EGFR) and G protein-coupled receptor (GPCR) signaling, and it directly binds calmodulin, actin, and F-actin. Through these interactions, ITPKA modulates the phosphorylation of Akt and the dynamics of filamentous actin, thereby influencing cellular migration, proliferation, and apoptotic signaling.
Within the HCT 116 colorectal carcinoma background, which harbors oncogenic KRAS and mutant TP53, ITPKA disruption offers a sophisticated model to examine how altered inositol phosphate metabolism intersects with dominant cancer drivers. KRAS-driven signaling via the MAPK and PI3K pathways engages calcium mobilization and actin remodeling, processes that are tightly controlled by ITPKA. Consequently, loss of ITPKA may impair directional migration, reduce proliferative capacity under growth factor stimulation, or alter the apoptotic balance, potentially sensitizing cells to targeted therapies. This model is exceptionally suited for mechanistic studies on calcium-dependent tumor cell motility, survival, and drug responsiveness.
Typical experimental applications include live-cell calcium imaging with fluorescent indicators to monitor spatiotemporal Ca2+ dynamics, Boyden chamber and wound-healing assays to quantify cell migration and invasion, and western blot analysis of Akt phosphorylation as a readout of downstream pathway activation. Apoptosis assays using annexin V or caspase-3/7 activation can assess survival phenotypes. The knockout cells also enable metabolic profiling of inositol phosphate species via HPLC or mass spectrometry, providing insights into how ITPKA loss re-routes phosphate metabolism. Drug resistance investigations benefit from the ability to couple ITPKA status with chemosensitivity profiling. For details or to request a quotation, please contact Ascent Research.
