ITGB4 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from HeLa cells, providing a mixed pool with disrupted ITGB4 gene function. This product allows researchers to directly assess the collective impact of integrin ??4 loss in an epithelial cancer background without single-cell cloning bias. By using a polyclonal knockout strategy, the model avoids potential artifacts from clonal selection and represents a practical tool for robust functional studies.
The parental HeLa line originates from human cervical adenocarcinoma and maintains integrated human papillomavirus type 18 (HPV18) sequences. These adherent epithelial cells are widely used to investigate tumor cell adhesion, migration, and signal transduction. HeLa cells express a broad array of integrin adhesion receptors, making them a suitable host for studying alpha6beta4 integrin-dependent processes. Their tractable growth characteristics and ease of transfection further facilitate downstream molecular analyses.
ITGB4 encodes the integrin ??4 subunit, which exclusively pairs with ITGA6 to form the laminin?332 receptor ??6??4. This complex is essential for hemidesmosome assembly, anchoring cells to basement membranes. Upon laminin?332 binding, ITGB4 recruits the adaptor protein SHC1 and the tyrosine kinase SRC, activating two major signaling cascades: PI3K/AKT and RAS/MAPK. Upstream, the receptor is regulated by the matrix ligand laminin?332 and by growth factors such as EGF and HGF, while transcription factors TP63 and AP?1 control ITGB4 expression. Downstream, the signals regulate AKT1, ERK1/2 (MAPK1/3), NF???B, STAT3, and the epithelial?mesenchymal transition (EMT) transcription factors SNAI1 and TWIST1. ITGB4 also interacts with plectin for cytoskeletal linkage and with receptor tyrosine kinases ErbB2 (HER2) and MET, integrating adhesion with growth factor signaling.
In the HeLa context, ITGB4 contributes to adhesion?dependent and ?independent signaling that promotes invasive behavior. Knockout of ITGB4 in this polyclonal population disrupts hemidesmosome assembly, reduces adhesion to laminin?332, and attenuates PI3K/AKT and MAPK pathway activation, thereby decreasing migratory and invasive capacity. This model is particularly relevant for dissecting how ??6??4 signaling intersects with HPV18 oncoprotein functions and for exploring integrin?mediated resistance mechanisms in cervical carcinoma.
Typical applications include Transwell migration and invasion assays, cell adhesion assays on laminin?332, and immunofluorescence to monitor hemidesmosome structure. Western blotting and RT?qPCR confirm ITGB4 depletion, while flow cytometry quantifies surface integrin loss. Co?immunoprecipitation and phospho?epitope analysis of AKT and ERK enable assessment of altered signaling. The cells are valuable for studying growth factor?Cintegrin crosstalk (EGF/HGF), drug sensitivity screening, and modeling metastatic progression in an isogenic background. For further information, please contact Ascent Research.
