The INPP5J Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the INPP5J gene, which encodes a phosphoinositide 5-phosphatase. This polyclonal knockout model eliminates INPP5J expression in a heterogeneous pool of HEK293T cells, enabling direct interrogation of INPP5J-dependent functions in PI3K/AKT signaling. The knockout population is generated via CRISPR/Cas9-mediated gene disruption, providing a physiologically relevant loss-of-function system without clonal selection artifacts.
INPP5J Knockout HEK293T Polyclonal Cells are derived from the widely used HEK293T host cell line, a human embryonic kidney epithelial line transformed with sheared adenovirus type 5 DNA and stably expressing the SV40 large T antigen. This genetic background confers high transfection efficiency and robust recombinant protein expression capacity, making HEK293T an ideal platform for mechanistic studies and large-scale functional genomics applications. The cells exhibit adherent growth and maintain typical epithelial morphology.
INPP5J functions as a lipid phosphatase that specifically hydrolyzes the D-5 phosphate from phosphatidylinositol (3,4,5)-trisphosphate (PIP3) and phosphatidylinositol (4,5)-bisphosphate (PI(4,5)P2). By depleting PIP3 pools at the plasma membrane, INPP5J negatively regulates PI3K/AKT signaling, attenuating AKT kinase activation and its downstream phosphorylation events. Key molecular connections include upstream activation by receptor tyrosine kinases (such as EGF and insulin receptors) and downstream modulation of AKT effectors like FOXO transcription factors, GSK-3??, mTORC1, BAD, and p27(Kip1). INPP5J also interacts with 14-3-3 proteins, cortactin, and the PI3K p85 regulatory subunit, integrating signals that control cell survival, proliferation, and cytoskeletal dynamics.
In HEK293T cells, the loss of INPP5J removes a critical brake on PIP3-dependent signaling, leading to sustained AKT phosphorylation and enhanced downstream pathway activity. This genetic perturbation is particularly informative given the HEK293T cell’s well-characterized signal transduction landscape and the presence of SV40 large T antigen, which inactivates p53 and Rb, creating a context that mimics certain oncogenic backgrounds. Consequently, the knockout model allows researchers to isolate the tumor-suppressive functions of INPP5J and assess its role in apoptosis regulation, cell cycle progression, and migratory behavior within an epithelial cell framework.
This knockout product supports a wide array of research applications aimed at deciphering PI3K/AKT pathway dysregulation in cancer and metabolic diseases. Users can employ the cells in Western blotting for phospho-AKT (Ser473), immunofluorescence-based PIP3 quantification, MTT proliferation assays, Transwell migration/invasion tests, and Annexin V apoptosis assays. The polyclonal knockout pool is also suited for transcriptomic analysis, RT-qPCR, and high-throughput drug screens against PI3K inhibitors. Typical disease models include glioblastoma, breast carcinoma, type 2 diabetes, and intellectual disability. For further technical details or customized support, please contact Ascent Research.
