IMPA1 Knockout A-549 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population of A-549 cells with targeted disruption of the IMPA1 gene. The polyclonal format comprises a heterogeneous mix of edits, ensuring a robust loss-of-function model without clonal selection. This product enables dissection of IMPA1-dependent pathways in a human lung adenocarcinoma background.
The A-549 cell line, derived from a 58-year-old Caucasian male with lung adenocarcinoma, serves as a model of type II alveolar epithelial carcinoma. Widely used in respiratory disease and oncology research, these cells provide a clinically relevant platform for studying cancer biology and drug responses. The IMPA1 knockout in this host allows investigation of inositol metabolism in non-small cell lung cancer.
IMPA1 hydrolyzes inositol monophosphate to myo-inositol, a precursor for phosphatidylinositol (PI) synthesis. It is inhibited by lithium and requires magnesium as a cofactor. IMPA1 activity sustains phosphatidylinositol-4,5-bisphosphate (PIP2) and phosphatidylinositol-3,4,5-trisphosphate (PIP3) pools, which activate Akt and protein kinase C (PKC) signaling. The enzyme can heterodimerize with IMPA2. Knockout of IMPA1 depletes myo-inositol, impairs PIP2-dependent cascades, and attenuates Akt/PKC pathway activation, leading to reduced proliferation and increased apoptosis. Downstream effects also involve IP3/DAG-mediated calcium signaling and GSK3?? modulation, linking inositol metabolism to Wnt and calcium pathways.
In A-549 cells, IMPA1 knockout compromises PI signaling, a pathway often dysregulated in cancer. Loss of IMPA1 reduces Akt-dependent survival signals, sensitizing cells to apoptotic stimuli. This model is particularly valuable for studying lithium’s anticancer mechanism, as lithium directly inhibits IMPA1. It also enables examination of cross-talk between inositol phosphate metabolism and Wnt/calcium pathways in epithelial tumor progression.
Research applications include quantifying myo-inositol by LC-MS, measuring phospho-Akt (Ser473) and PIP2 by ELISA, and performing MTT proliferation, annexin V apoptosis, and Transwell migration/invasion assays. The cells support lithium dose-response studies and inhibitor screening. This tool aids in modeling bipolar disorder-related signaling and elucidating PIP2-dependent mechanisms in lung adenocarcinoma. For technical inquiries, contact Ascent Research.
