The INPPL1 Knockout HT29 Polyclonal Cells represent a CRISPR/Cas9-edited cell population in which the INPPL1 gene has been disrupted via targeted genome editing. As a polyclonal knockout pool, this product contains a heterogeneous mixture of edited HT29 cells, each carrying targeted disruption of the INPPL1 gene. This format provides a robust and ready-to-use model for studying the consequences of INPPL1 deficiency without the clonal selection biases inherent in single-cell-derived lines.
The host cell line, HT29, is a well-characterized human colorectal adenocarcinoma line derived from a primary tumor and exhibits epithelial morphology. Under appropriate culture conditions, HT29 cells can differentiate into enterocyte-like cells, recapitulating aspects of intestinal epithelial biology. The cells endogenously express core components of the PI3K/AKT and insulin signaling cascades, including insulin receptors, PI3K, PTEN, and AKT, establishing a physiologically relevant context for examining INPPL1 function.
INPPL1 encodes SHIP2, a phosphatase that specifically hydrolyzes the 5-phosphate of phosphatidylinositol 3,4,5-trisphosphate (PIP3) to generate PI(3,4)P2, thereby attenuating the PI3K/AKT signaling axis. Its enzymatic activity is stimulated by upstream regulators such as growth factors (EGF, PDGF), insulin, integrin receptors, and SRC kinases. SHIP2 interacts with a network of signaling mediators including the adaptor proteins SHC, IRS1, and p130Cas, and recruits filamin A and c-Cbl. By limiting PIP3 availability, INPPL1 restrains activation of downstream effectors including AKT, GSK3??, mTOR, FOXO transcription factors, and the small GTPase Rac1, and its disruption promotes sustained pathway activation.
In the colorectal cancer context of HT29 cells, INPPL1 knockout removes a critical negative regulator of PI3K-dependent growth and survival signals. Loss of SHIP2 activity leads to accumulation of PIP3 and hyperactivation of AKT, which in turn phosphorylates and inhibits GSK3??, activates mTOR, and excludes FOXO factors from the nucleus??events that collectively drive enhanced cell cycle progression, resistance to apoptosis, and increased migratory capacity. This model therefore recapitulates oncogenic signaling perturbations frequently observed in colorectal tumors harboring dysregulated PI3K pathway activity, making it a valuable tool for exploring SHIP2??s tumor-suppressive functions.
Researchers can employ this INPPL1 polyclonal knockout pool in a spectrum of functional assays, including insulin stimulation experiments to monitor AKT phosphorylation dynamics by western blotting or flow cytometry, cell proliferation and wound healing migration assays to assess invasiveness, and RT-qPCR to quantify changes in FOXO target genes. The model is also suited for pharmacological testing of PI3K or mTOR inhibitors under loss-of-function conditions and for studying crosstalk between insulin and oncogenic pathways. For further information or to inquire about this product, please contact Ascent Research.
