The ITPKA Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population targeting the ITPKA gene. This heterogeneous pool provides a loss-of-function model for studying inositol-trisphosphate 3-kinase A (ITPKA) functions without clonal selection. The polyclonal format captures diverse editing events, making it ideal for population-level signaling and functional genomics studies.
The parental HEK293T cell line is a human embryonic kidney epithelial derivative that stably expresses SV40 large T-antigen, enabling episomal replication of plasmids with an SV40 origin. This feature supports high-efficiency transient transfection and viral packaging, establishing HEK293T as a versatile tool in molecular biology for protein expression, lentiviral production, and biochemical assays.
ITPKA is a key regulator of inositol phosphate signaling and actin dynamics. It phosphorylates IP3 to IP4, dampening IP3-mediated calcium release from intracellular stores. ITPKA is activated by GPCR/PLC signaling and CaMKII, and it interacts with calmodulin, F-actin, 14-3-3??, and the IP3 receptor. Beyond catalysis, ITPKA directly bundles F-actin, influencing actin filament organization and cell migration through cofilin pathways. Thus, ITPKA integrates calcium and cytoskeletal signaling.
In HEK293T cells, ITPKA knockout permits dissection of its dual roles in calcium flux and actin remodeling. Despite their epithelial origin, HEK293T cells display actin-driven processes and respond to GPCR agonists, making them a useful model for studying migration-related mechanisms. Loss of ITPKA enables examination of how disrupted IP3 metabolism and actin bundling alter cellular responses to stimuli, with relevance to cancer metastasis and neuronal development.
This knockout pool supports diverse assays: western blotting and RT-qPCR for gene expression validation; calcium imaging for altered signaling dynamics; phalloidin staining for actin cytoskeleton visualization; and migration/invasion assays to assess phenotypic effects. Co-immunoprecipitation and phospho-signaling analysis map interaction networks. RNA-seq may reveal transcriptomic changes. For additional information, please contact Ascent Research.
