INPPL1 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HEK293T human cell line, engineered to disrupt the INPPL1 gene locus. This polyclonal pool, generated through non-clonal targeting, provides a physiologically relevant loss-of-function model that preserves the inherent heterogeneity of the parental population. The product enables robust investigation of SHIP2-dependent signaling without the selective pressure of single-cell cloning, making it particularly suitable for pathway analysis in a diverse cellular context.
HEK293T cells are a widely utilized derivative of the HEK293 line, originally transformed by sheared adenovirus 5 DNA and further modified to stably express the SV40 large T antigen. This human embryonic kidney epithelial cell line supports high-level transient protein expression and efficient lentivirus production, and serves as a foundational tool for studying signal transduction, protein interactions, and cellular proliferation. Its well-characterized and easily transfectable nature makes it an ideal host for generating gene-edited knockout models.
INPPL1 encodes SHIP2, a 5??-inositol phosphatase that hydrolyzes the lipid second messenger PIP3 to PI(3,4)P2, thereby attenuating PI3K/AKT signaling. SHIP2 functions downstream of receptors such as INSR and EGFR, and interacts with adaptor proteins including SHC1, GRB2, and IRS1. By downregulating PIP3 levels, SHIP2 negatively regulates key effectors such as AKT1, mTORC1, GSK3??, and FOXO transcription factors, while also modulating Rac1-mediated actin dynamics. Its activity is tightly linked to insulin sensitivity, growth control, and cytoskeletal organization.
Disruption of INPPL1 in HEK293T cells results in sustained PIP3 accumulation, leading to constitutive activation of AKT and downstream mTORC1. In this transformed background, loss of SHIP2 further amplifies PI3K-dependent growth and proliferation while potentially impairing negative feedback loops. The polyclonal nature of the knockout pool allows assessment of population-level signaling alterations and functional consequences, including changes in cell cycle progression and migratory behavior, without the confounding effects of clonal selection.
This polyclonal knockout model is well-suited for mechanistic studies of PI3K/AKT pathway hyperactivation, insulin and growth factor signaling, and oncogenic processes. Applications include drug screening for AKT or mTOR inhibitors, analysis of cell migration via wound healing assays, and quantification of PIP3 levels by ELISA. The cells can be used in western blotting for phospho-AKT (Ser473) and phospho-S6, RT-qPCR for FOXO target genes, and co-immunoprecipitation experiments to probe altered protein complexes. For further technical information or assistance with assay design, please contact Ascent Research.
