The INPPL1 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function studies of the INPPL1 gene in a human haploid background. This product consists of a heterogeneous pool of HAP1 cells carrying targeted disruptions of the INPPL1 locus, enabling investigation of SHIP2 phosphatase function without clonal selection.
HAP1 is a near-haploid human hematopoietic cell line derived from the KBM-7 chronic myeloid leukemia isolate. Its haploid karyotype reduces genetic redundancy, facilitating unambiguous genotype-phenotype correlations and making it a powerful platform for functional genomics, drug screening, and signaling pathway dissection.
INPPL1 encodes SHIP2, an inositol polyphosphate 5-phosphatase that hydrolyzes phosphatidylinositol (3,4,5)-trisphosphate (PIP3) to phosphatidylinositol (3,4)-bisphosphate (PI(3,4)P2), thereby attenuating PI3K/AKT signaling downstream of receptor tyrosine kinases (RTKs). SHIP2 is activated by insulin, IGF-1, and EGF stimulation, and acts as a critical negative regulator of the PI3K/AKT pathway. Its activity reduces AKT phosphorylation and subsequent activation of downstream effectors including mTORC1, GSK3??, and FOXO transcription factors, while impacting small GTPases Rac and Rho and glucose transporter GLUT4 translocation. SHIP2 interacts with adaptor proteins such as GRB2, SHC, and IRS1, and the p85 regulatory subunit of PI3K, integrating signals at the plasma membrane.
In the HAP1 leukemia background, loss of INPPL1 is anticipated to sustain PI3K/AKT activation, potentially enhancing cell proliferation, survival, and metabolic reprogramming. This model is particularly relevant for exploring mechanisms underlying insulin resistance, oncogenic transformation, and metastasis, as SHIP2 has been implicated in type 2 diabetes, obesity, and various cancers. The near-haploid nature of HAP1 cells ensures that gene disruption generates a functionally null state without compensation from a second allele, providing a clean loss-of-function system for studying SHIP2-dependent phenotypes.
Researchers can employ these polyclonal knockout cells to dissect insulin signaling kinetics via phospho-AKT western blot or flow cytometry, quantify glucose uptake in response to insulin, assess cell migration using scratch or transwell assays, or screen chemical libraries for SHIP2 inhibitors. They also serve as a valuable control for rescue experiments reintroducing wild-type or mutant SHIP2. Typical applications include modeling type 2 diabetes, investigating oncogenic PI3K/AKT pathway addiction, and validating SHIP2 as a therapeutic target. For further details or customization options, please contact Ascent Research.
