The ILKAP Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the widely used HEK293T human embryonic kidney cell line. This product provides a genetically disrupted ILKAP gene, enabling researchers to study the loss-of-function effects of this critical serine/threonine phosphatase. The polyclonal nature of the knockout population preserves the inherent heterogeneity of CRISPR-mediated gene disruption, making it suitable for bulk population studies without the need for clonal isolation. This model is ideal for investigating the role of ILKAP in integrin signaling and associated pathways in a high-throughput, tractable cellular background.
The HEK293T host cell line is a derivative of the HEK293 human embryonic kidney cell line, engineered to stably express the SV40 large T antigen, which facilitates episomal replication of plasmids containing the SV40 origin of replication. This feature renders HEK293T cells exceptionally efficient for transient protein expression, making them a cornerstone model for biochemical and cell biological studies. Widely employed for signaling pathway dissection, protein?Cprotein interaction analysis, and functional genomics, HEK293T cells offer a robust platform for exploring the consequences of targeted gene disruption. Their epithelial origin and expression of key integrin subunits further support the study of adhesion-dependent signaling events, providing a relevant context for ILKAP functional analysis.
ILKAP encodes a serine/threonine phosphatase that functions as a negative regulator of integrin-linked kinase (ILK) signaling, thereby modulating critical cellular processes including adhesion, migration, and survival. Mechanistically, ILKAP directly dephosphorylates ILK at Ser343, suppressing ILK kinase activity and attenuating downstream Akt/GSK3?? signaling cascades. This phosphatase is activated by upstream cues such as integrin engagement, growth factor stimulation, cellular stress, and TGF-??, and it forms regulatory complexes with ILK, PINCH, Parvin, and PP2A regulatory subunits. Within the broader network, ILKAP intersects with integrin?CFAK?CILK?CAkt/GSK3?? and Wnt/??-catenin pathways, thereby influencing key transcriptional and cytoskeletal programs. Disruption of ILKAP thus derepresses ILK-dependent signaling, providing a direct model to interrogate these interconnected axes.
In the HEK293T cellular context, ILKAP knockout has particular significance for dissecting the molecular underpinnings of tumor suppression and cell motility. HEK293T cells exhibit robust integrin-mediated adhesion and are amenable to manipulation of downstream effectors such as Akt and GSK3??. Loss of ILKAP function is expected to enhance phosphorylation of ILK and its substrates, mimicking oncogenic signaling states observed in cancers and fibrotic diseases. This model is well-suited to study how aberrant ILK activation drives pro-survival, migratory, or hypertrophic phenotypes, and to evaluate the interplay between ILKAP and TGF-??-induced responses. By combining gene disruption with the biochemical tractability of HEK293T cells, researchers can dissect the precise contribution of ILKAP to cellular homeostasis and disease progression.
The ILKAP Knockout HEK293T Polyclonal Cells enable a broad range of advanced research applications. They are ideally used for quantitative analysis of ILK phosphorylation status via phospho-specific western blotting, RT-qPCR profiling of downstream gene expression changes, and phospho-Akt analysis to monitor pathway activity. Functional assays such as wound-healing migration assays, adhesion assays, and live-cell imaging can reveal phenotypic consequences of ILKAP loss. Co-immunoprecipitation experiments facilitate mapping of altered ILK interaction networks, while RNA-sequencing provides global transcriptional insights. These cells support investigations into tumor suppressor mechanisms, cardiac hypertrophy signaling, and fibrogenesis, and they serve as a valuable tool for target validation and drug screening in integrin-dependent diseases. For further product details, please contact Ascent Research.
