The ILK Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in which the ILK gene has been disrupted to abolish integrin-linked kinase expression. This loss-of-function model enables robust interrogation of ILK-dependent signaling and adhesive functions without clonal selection, providing a genetically diverse yet ILK-deficient cellular pool for population-level studies.
HAP1 is a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia line. Its haploid karyotype allows unambiguous genotype-phenotype correlations, as a single gene disruption eliminates functional expression without interference from a second allele. This makes HAP1 particularly valuable for functional genomics screens, drug target validation, and mechanistic studies in a genetically stable background.
ILK functions as a serine/threonine kinase and scaffold at focal adhesions, linking integrin receptors to the actin cytoskeleton. It forms the IPP complex with PINCH (particularly PINCH1) and parvin (??, ??, ??). ILK transduces extracellular matrix signals by directly phosphorylating AKT and GSK-3??, thereby promoting cell survival and proliferation, and stabilizes ??-catenin to activate TCF/LEF-mediated transcription, regulating cell fate and migration. Upstream, ILK is activated by integrin clustering, PI3K, and growth factor receptors such as EGFR and IGF-1R, with PTEN providing negative regulation. Key downstream effectors include paxillin, FAK, and Rho GTPases, which orchestrate cytoskeletal dynamics.
In the haploid HAP1 background, ILK knockout yields complete loss of function, enabling clear dissection of ILK-dependent processes. Disruption of the IPP complex impairs integrin-to-cytoskeleton signal transduction, making this model particularly suitable for studying focal adhesion dynamics, cell adhesion, and migration. It also allows systematic analysis of ILK crosstalk with PI3K/AKT and Wnt/??-catenin pathways, as well as the scaffolding versus kinase functions of ILK.
This knockout cell pool is well-suited for experiments such as western blotting to confirm ILK loss and monitor AKT/GSK-3?? phosphorylation, immunofluorescence for focal adhesion proteins, co-immunoprecipitation to assess IPP complex assembly, and RT-qPCR for ??-catenin target genes. Functional assays including cell adhesion, migration/invasion, and apoptosis can elucidate ILK??s role in cancer cell behavior. Drug sensitivity testing with PI3K or AKT inhibitors helps explore ILK-mediated chemoresistance. For more information, please contact Ascent Research.
