INPP5K Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human Jurkat T-lymphocyte cell line. This product provides a loss-of-function model for the INPP5K gene, which encodes inositol polyphosphate-5-phosphatase SKIP. The polyclonal format comprises a heterogeneous pool of gene-edited cells, enabling robust study of INPP5K deficiency without the bottlenecks of single-cell cloning. CRISPR/Cas9-mediated gene disruption has been applied to ablate INPP5K expression, yielding a versatile tool for dissecting phosphoinositide signaling networks.
Jurkat cells are an immortalized CD4+ T-cell leukemia line originally established from a 14-year-old male with acute lymphoblastic leukemia. They serve as a widely used model for T-cell receptor (TCR) signaling, activation, and apoptosis. Their rapid proliferation and well-characterized signaling pathways make them ideal hosts for investigating the roles of immunomodulatory genes. The CD4+ T-cell context is particularly relevant for studying PI3K/AKT/mTOR axis regulation and its impact on T-cell fate decisions.
INPP5K functions as a phosphoinositide phosphatase that catalyzes the hydrolysis of phosphatidylinositol (3,4,5)-trisphosphate (PIP3) and inositol (1,4,5)-trisphosphate (IP3), thereby terminating downstream signaling through the PI3K/AKT pathway. It operates downstream of receptor tyrosine kinases such as insulin receptor, IGF-1 receptor, and EGF receptor, and upstream of AKT, GSK3, mTOR, FOXO, and NF-??B. INPP5K also interacts with Filamin A, linking phosphoinositide metabolism to actin cytoskeleton remodeling. Consequently, INPP5K knockout in Jurkat cells results in elevated PIP3 levels, constitutive AKT phosphorylation at serine 473, and hyperactivation of mTOR signaling, alongside potential cytoskeletal alterations due to disrupted Filamin A binding.
In the Jurkat T-cell model, INPP5K loss intensifies TCR-dependent and -independent PI3K/AKT/mTOR signaling, leading to enhanced cell proliferation, survival, and cytokine production (e.g., IL-2). This hyperactivation may mimic oncogenic conditions observed in T-cell leukemias and other cancers, while also impacting insulin-dependent metabolic pathways. The interplay between elevated PIP3, AKT hyperphosphorylation, and transcriptional regulators like FOXO and NF-??B provides a platform to study T-cell activation checkpoints and resistance mechanisms. Furthermore, the role of INPP5K in cytoskeletal organization suggests utility in examining T-cell migration and adhesion, processes critical in immune surveillance and metastasis.
Applications include western blotting for phospho-AKT (S473), PIP3 mass ELISA, flow cytometry for CD69/CD25, CFSE proliferation assays, apoptosis analysis, and RT-qPCR for IL-2 and AKT targets. Co-immunoprecipitation can assess INPP5K interactors. These cells support studies from T-cell signaling to cancer metabolism and immune checkpoint research. For further details, please contact Ascent Research.
