The ITGB4 Knockout HEK293T Polyclonal Cells product provides a polyclonal population of HEK293T cells harboring targeted disruption of the ITGB4 gene via CRISPR/Cas9-mediated gene editing. This knockout model enables loss-of-function studies of integrin beta-4 (ITGB4), a critical transmembrane receptor subunit that pairs with integrin alpha-6 to form the laminin-binding ??6??4 integrin heterodimer. The polyclonal format preserves a heterogeneous knockout pool, reflecting diverse editing outcomes across the cell population, thereby offering a robust system for investigating ITGB4-dependent biological processes without clonal selection biases.
The host cell line, HEK293T, is an epithelial derivative of human embryonic kidney that stably expresses the SV40 large T antigen, which enhances plasmid replication and transient protein expression. Its epithelial morphology and high transfection efficiency make it a preferred system for exploring integrin-mediated adhesion and signaling events central to epithelial cell biology.
Integrin beta-4 (ITGB4) specifically partners with integrin alpha-6 (ITGA6) to form the ??6??4 integrin, the primary receptor for laminin-332 in epithelial basement membranes. The ??6??4 integrin is a core structural component of hemidesmosomes, where it connects extracellular matrix to the intermediate filament cytoskeleton through direct interactions with plectin and BPAG1/BPAG2. Laminin-332 engagement triggers phosphorylation of downstream kinases FAK and Src, which activate the PI3K-Akt axis to promote cell survival and proliferation. Additionally, ITGB4 functionally cooperates with receptor tyrosine kinases EGFR and ErbB2, influencing Rho GTPase-mediated cytoskeletal remodeling and STAT3-driven transcription. ITGB4 expression is transcriptionally regulated by p63 and post-transcriptionally modulated by miR-205, tying its abundance to epithelial differentiation and oncogenic processes.
In HEK293T cells, ITGB4 knockout ablates ??6??4 integrin function, eliminating laminin-332 binding and disrupting adhesion structures. This model enables dissection of ITGB4-specific contributions to cell-matrix adhesion, migration, and signaling without interference from other integrins. The polyclonal knockout pool reflects heterogeneous loss-of-function, advantageous for studying phenotypes linked to ITGB4 dysregulation in cancers such as squamous cell carcinoma. It also facilitates rescue experiments for structure-function analysis of ITGB4 domains.
Key applications include adhesion and migration assays, laminin-332 binding studies, immunofluorescence analysis of hemidesmosome components, Western blotting and phospho-signaling profiling of FAK/Src/Akt and STAT3 pathways, RT-qPCR analysis of gene regulation by p63 and miR-205, and cancer metastasis screens. The polyclonal cells are suitable for pooled functional genomics and biochemical studies. For additional information, contact Ascent Research.
