The IMPA2 Knockout HEK293T Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal knockout population of HEK293T cells with targeted disruption of the IMPA2 gene. The polyclonal format offers a pooled loss-of-function model that minimizes clonal artifacts and is ideal for bulk functional analyses.
The HEK293T cell line is a human embryonic kidney epithelial line stably expressing the SV40 large T-antigen, which permits episomal replication of plasmids containing an SV40 origin. Derived from HEK293 cells transformed with adenovirus 5 DNA, HEK293T cells are widely used for transient transfection and viral production due to their high transfection efficiency and robust growth.
IMPA2 encodes inositol monophosphatase 2, which catalyzes the hydrolysis of inositol monophosphate to free inositol, a key step in phosphoinositide recycling. This enzyme functions downstream of phospholipase C (PLC)-mediated cleavage of PI(4,5)P2 into IP3 and DAG, and its activity is inhibited by lithium. By forming homodimers, IMPA2 sustains cellular inositol pools required for phosphoinositide-dependent signaling, including calcium mobilization. Its disruption therefore mimics lithium-induced inositol depletion and impairs IP3-mediated calcium release.
In HEK293T cells, IMPA2 knockout eliminates endogenous inositol monophosphatase activity, reducing free inositol and disrupting phosphoinositide recycling. This genetic model of inositol depletion alters PLC-dependent calcium signaling and phospho-signaling networks, providing a tool to study lithium’s molecular effects without pharmacological intervention. The HEK293T background retains functional G protein-coupled receptor signaling and calcium-release machinery, making it suitable for investigating inositol-dependent processes and neuronal-relevant pathways in an epithelial context.
Applications include mechanistic studies of lithium action, investigation of bipolar disorder pathophysiology, and screening of IMPA2 inhibitors. Typical assays encompass inositol monophosphatase activity measurements, LC-MS inositol quantification, calcium imaging with Fluo-4, cell proliferation under lithium, and phospho-signaling analyses such as pAkt. Transcriptomic profiling via RNA-seq further enables mapping of downstream gene expression changes. For additional information, please contact Ascent Research.
