This product comprises a CRISPR/Cas9-edited polyclonal population of SK-HEP-1 cells carrying a targeted disruption of the INPP5B gene. The cell pool provides a loss-of-function model for investigating inositol polyphosphate-5-phosphatase function, circumventing clonal artifacts through heterogeneous editing outcomes. The polyclonal format captures a range of allelic variants within the same culture, enabling robust assessment of INPP5B-dependent phenotypes under physiologically relevant expression variations.
The host cell line SK-HEP-1 originates from a human male hepatic adenocarcinoma and is widely established as an in vitro model for liver cancer research. It retains key epithelial and tumorigenic features, including anchorage-independent growth and modulation of phosphoinositide-dependent signaling pathways. This background is particularly suited for dissecting the contribution of lipid phosphatases to hepatocellular carcinoma progression and therapeutic resistance.
INPP5B encodes an inositol polyphosphate-5-phosphatase that preferentially dephosphorylates the 5-position of phosphatidylinositol (3,4,5)-trisphosphate (PIP3) to generate phosphatidylinositol (4,5)-bisphosphate (PIP2). By counteracting PI3K activity, INPP5B attenuates downstream AKT1 recruitment and activation, thereby regulating cell survival, proliferation, and cytoskeletal dynamics. Its function is positioned downstream of growth factor receptors such as EGFR and insulin receptor, and upstream of RAC1-dependent actin polymerization. Additionally, INPP5B interacts with endocytic regulators including ARF6, RAB5A, and APPL1, and shares functional domains with OCRL, linking it to clathrin-mediated endocytosis and focal adhesion dynamics.
In SK-HEP-1 cells, disruption of INPP5B is expected to elevate PIP3 levels, resulting in hyperactive AKT/mTOR signaling and concomitant alterations in actin filament organization and endocytic trafficking. These changes recapitulate oncogenic phosphoinositide dysregulation frequently observed in liver cancer, making the knockout model a powerful tool for examining how aberrant PIP3 metabolism drives malignant phenotypes such as enhanced migration and proliferation. Furthermore, the genetic interaction network with PTEN and PIK3CA offers a platform to dissect pathway cross-talk and assess compensatory mechanisms.
Typical applications include western blotting for phospho-AKT (Ser473), immunofluorescence for F-actin, PIP3 ELISA, scratch wound migration, MTS proliferation, and transferrin uptake endocytosis assays. This model supports drug sensitivity profiling against PI3K/AKT/mTOR, and dissection of INPP5B roles in hepatocellular carcinoma, renal carcinoma, and Lowe syndrome spectrum. Please contact Ascent Research for further technical details.
