The INPP5J Knockout HeLa Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout population derived from the HeLa human cervical epithelial cell line. This product provides a loss-of-function model for the INPP5J gene, enabling investigations into phosphoinositide metabolism and tumor suppressor mechanisms. The heterozygous polyclonal population carries diverse gene disruptions across the cell pool, offering a robust system for studying gene function without clonal isolation artifacts. These cells are suitable for assays requiring pooled knockout phenotypes, including signaling pathway analysis and functional genomics screens.
The HeLa cell line is an immortalized human cervical adenocarcinoma line that harbors human papillomavirus type 18 (HPV-18) sequences. As one of the most extensively used models in biomedical research, HeLa cells offer well-characterized growth properties, high transfection efficiency, and reproducible experimental conditions. Their epithelial origin and transformed nature make them particularly relevant for cancer biology studies, including investigations into tumor cell migration, invasion, and metastatic progression. The consistent genetic background of HeLa cells provides a reliable platform for assessing the consequences of INPP5J disruption on cellular behavior.
INPP5J encodes a phosphoinositide 5-phosphatase that specifically hydrolyzes the signaling lipids phosphatidylinositol 4,5-bisphosphate (PIP2) and inositol 1,4,5-trisphosphate (IP3). This enzymatic activity negatively regulates the PI3K/Akt signaling axis by reducing the availability of PIP2, thereby attenuating downstream Akt phosphorylation and activation. INPP5J functions downstream of receptor tyrosine kinases such as EGFR and PI3K, and its activity is transcriptionally regulated by the p53 tumor suppressor in response to DNA damage. The phosphatase directly impacts actin cytoskeleton dynamics by modulating Rac1 and Cofilin, which are critical for cell migration. By integrating signals from p53 and the extracellular matrix, INPP5J serves as a gatekeeper that restrains cytoskeletal reorganization and cell motility.
In the HeLa cervical cancer context, loss of INPP5J disrupts this regulatory checkpoint, leading to sustained PIP2 levels, hyperactivation of Akt signaling, and enhanced actin polymerization. This molecular profile promotes a more migratory and invasive phenotype, mirroring features of metastatic progression. The knockout model thus provides a valuable tool for dissecting the roles of phosphoinositide phosphatases in tumor suppression and for evaluating how HPV-induced oncogenic pathways intersect with phospholipid metabolism. Researchers can exploit this system to identify druggable nodes in the PI3K-Akt-Rac1 axis that contribute to cancer cell dissemination.
Typical applications include quantitative Western blotting for INPP5J, total Akt, and phospho-Akt to confirm pathway modulation; transwell migration and Matrigel invasion assays to measure metastatic potential; immunofluorescence staining to visualize actin stress fibers and focal adhesions; and proliferation studies under various growth conditions. These polyclonal knockout cells are also suitable for phospho-signaling arrays and drug sensitivity screens targeting PI3K or downstream effectors. For additional technical specifications or ordering information, please contact Ascent Research.
