The ITPKA Knockout A-549 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population in which the ITPKA gene has been disrupted. This product provides a heterogeneous pool of cells harboring various loss-of-function mutations, enabling the study of ITPKA deficiency within a lung adenocarcinoma epithelial background. As a polyclonal knockout population, it reflects the aggregate effects of diverse editing events, offering a robust model for functional genomics and drug target validation without requiring clonal isolation.
The A-549 host cell line was originally derived from the lung adenocarcinoma tissue of a 58-year-old Caucasian male. These adherent epithelial cells are a classic model for non-small cell lung cancer (NSCLC) research, exhibiting well-characterized growth properties and genetic features. The A-549 line retains key oncogenic signaling pathways and is widely employed for investigating tumor biology, metastatic mechanisms, and therapeutic responses.
ITPKA encodes an inositol-trisphosphate 3-kinase that catalyzes the phosphorylation of inositol 1,4,5-trisphosphate (IP3) to inositol 1,3,4,5-tetrakisphosphate (IP4). This enzymatic activity attenuates IP3-mediated calcium release from the endoplasmic reticulum by reducing IP3 levels and generating IP4, which may itself modulate calcium homeostasis. ITPKA also binds to filamentous actin (F-actin) through actin-binding motifs, directly influencing actin cytoskeleton organization. ITPKA is positioned downstream of EGF receptor signaling, HIF1A, and TP53, and it interacts with calmodulin in a calcium-dependent manner, as well as with the IP3 receptors ITPR1 and ITPR2. Through its dual actions on calcium flux and actin dynamics, ITPKA regulates downstream effectors such as NFAT signaling and IP4-sensitive calcium pools, thereby impacting cellular responses including proliferation and apoptosis.
In the context of A-549 lung adenocarcinoma cells, ITPKA knockout is particularly relevant for dissecting pathways driving tumor aggressiveness. The gene has been implicated in promoting migration and invasion, processes central to metastasis, partly via its F-actin binding activity. Moreover, altered inositol phosphate signaling and calcium oscillations can influence apoptotic thresholds and cell cycle progression. This knockout model allows researchers to systematically examine how ITPKA deficiency affects metastatic potential, cytoskeletal remodeling, and calcium-dependent signaling in a NSCLC-derived line, providing insights into solid tumor biology.
The ITPKA Knockout A-549 Polyclonal Cells are suited for a broad range of experimental applications. They can be utilized in Transwell migration and invasion assays to quantify changes in motility, in calcium imaging studies to measure IP3-dependent store release, and in immunofluorescence experiments to visualize actin cytoskeleton alterations. Co-immunoprecipitation with antibodies against ITPR1, calmodulin, or F-actin can map interaction networks, while downstream readouts via RT-qPCR and Western blotting validate pathway perturbations. The model also supports drug discovery efforts targeting ITPKA-related signaling and apoptosis pathway analyses using Annexin V/PI staining. For further details or specific inquiry, please contact Ascent Research.
