The ILKAP Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HAP1 human near-haploid cell line, engineered to disrupt the ILKAP gene. This polyclonal pool offers a loss-of-function model for studying ILKAP??s role in integrin-mediated signal transduction, without the clonal selection bottlenecks inherent in monoclonal derivatives. The heterogeneous knockout population preserves genetic diversity while broadly ablating ILKAP expression, making it suitable for pooled functional genomics and signaling studies.
HAP1 cells originate from the KBM-7 chronic myeloid leukemia (CML) line and possess a near-haploid karyotype, which simplifies gene editing and genetic manipulation. These adherent cells retain many signaling characteristics of hematopoietic lineages and are widely employed in genome-wide knockout screens, drug sensitivity profiling, and pathway interrogation. Their haploid state reduces the complexity of interpreting loss-of-function phenotypes, thereby facilitating robust analysis of signaling networks where ILKAP functions as a negative regulator.
ILKAP encodes a serine/threonine protein phosphatase that directly dephosphorylates integrin-linked kinase (ILK) on regulatory residues, thereby decreasing ILK kinase activity. This dephosphorylation attenuates downstream AKT phosphorylation at Thr308 and Ser473, reducing AKT-mediated signaling that drives cell proliferation, migration, and survival. ILKAP operates within the integrin adhesion complex, where it interacts with ILK, PINCH, and parvin to modulate signaling downstream of integrin ??1 engagement. Consequently, ILKAP loss unleashes constitutive ILK?CAKT signaling, leading to enhanced phosphorylation of GSK3?? and stabilization of ??-catenin, thus potentiating Wnt pathway crosstalk. Upstream, ILKAP activity is stimulated by integrin engagement and cell?Cmatrix adhesion, positioning it as a critical feedback brake on mechanical and growth factor signals.
In the HAP1 CML background, where ILK?CAKT signaling fosters leukemogenesis, ILKAP disruption provides a disease-relevant model to examine how loss of phospho-regulation enhances tumor cell survival and drug resistance. The polyclonal knockout population allows systematic dissection of ILKAP??s role in adhesion-dependent signaling without clonal artifacts, enabling studies of pathway rewiring under endogenously titrated integrin signals. This model is particularly valuable for probing the interplay between integrin signaling and oncogenic kinase networks, given that HAP1 cells retain BCR?ABL expression.
Typical applications include monitoring phospho-AKT (Ser473) and phospho-GSK3?? (Ser9) levels by western blotting, performing cell adhesion and migration assays under various matrix-coated conditions, and conducting proliferation assays to assess sensitivity to kinase inhibitors. Co-immunoprecipitation of the ILK complex components can be employed to investigate how ILKAP loss alters complex assembly. Furthermore, these cells serve as a powerful tool in drug sensitivity screens aimed at identifying compounds that exploit ILK?CAKT pathway addiction. For additional technical information, custom orders, or bulk pricing, please contact Ascent Research.
