The ITGA3 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the HT29 human colorectal adenocarcinoma cell line, in which ITGA3 gene expression is disrupted to create a loss-of-function model for studying integrin alpha-3 biology. This heterogeneous pool preserves the parental line??s diversity and enables robust pooled assays, avoiding clonal selection artifacts. CRISPR/Cas9-mediated gene disruption abrogates alpha-3 subunit production, impairing alpha3beta1 heterodimer formation.
HT29 cells originate from a 44-year-old Caucasian female with colorectal adenocarcinoma and are tumorigenic in nude mice. These epithelial cells retain intestinal characteristics, including the capacity to differentiate and secrete mucus, and are widely used in cancer research, drug transport, and barrier function studies. Their relevance to colorectal pathology makes them a suitable host for interrogating ITGA3-dependent mechanisms in a disease-relevant context.
ITGA3 encodes the integrin alpha-3 subunit, which partners with ITGB1 to form the alpha3beta1 receptor, binding laminin-332, collagen IV, and fibronectin. Ligand engagement activates FAK and Src, driving PI3K-AKT and MAPK/ERK cascades that regulate adhesion, survival, and migration. ITGA3 expression is controlled by TGF-beta1, SP1, AP-1, EGF, and HIF-1??, and its signaling converges on effectors including RhoA, Rac1, MMP9, Bcl-2, and cyclin D1. ITGA3 also interacts with CD151, EGFR, and c-Met, integrating extracellular cues into invasive programs. Knockout in this model eliminates alpha3beta1-mediated adhesion and downstream phosphorylation events.
In the HT29 adenocarcinoma background, ITGA3 loss impairs laminin-dependent adhesion, migration, and invasion, offering a system to dissect its role in metastasis and drug resistance. Abrogation of FAK/Akt survival signaling sensitizes cells to anoikis, while disruption of integrin?Cgrowth factor crosstalk may alter chemosensitivity. This model enables examination of compensatory changes in tetraspanin networks and matrix metalloproteinase activity, illuminating tumor adaptation.
Applications include adhesion assays on laminin/collagen, Boyden chamber migration/invasion, Western blotting for ITGA3, phospho-FAK, and ERK1/2, RT-qPCR, flow cytometry, immunofluorescence, and co-immunoprecipitation of integrin complexes. The model is valuable for integrin-targeted drug screening, intestinal barrier research, and ECM signaling studies. For further information or technical support, please contact Ascent Research.
