The ITGA2 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from HT29 human colorectal adenocarcinoma cells, carrying a targeted disruption of the ITGA2 gene. This pool enables loss-of-function studies of integrin ??2, which heterodimerizes with integrin ??1 to form the ??2??1 collagen/laminin receptor. The polyclonal composition provides a diverse knockout spectrum, avoiding clonal selection biases and offering a robust model for studying integrin-mediated processes.
The HT29 parental line is a human colorectal adenocarcinoma epithelial model isolated from a primary tumor of a 44-year-old Caucasian female. Widely used in intestinal biology, colorectal cancer, and drug transport research, HT29 cells retain the ability to differentiate into enterocyte-like phenotypes. This background offers a physiologically relevant system for examining adhesion receptor roles in malignant progression, metastasis, and therapy resistance.
ITGA2 encodes integrin ??2, which partners with ITGB1 to form the ??2??1 heterodimer, a receptor for collagen types I and IV, laminin, and fibronectin. Ligand binding triggers FAK autophosphorylation at Y397, recruiting Src kinase to phosphorylate adaptors p130Cas and Crk, leading to activation of the DOCK180?CRac1 axis and RhoA/Cdc42 GTPases. Concurrently, MAPK signaling proceeds via GRB2?CSOS?CRas?CRaf?CMEK?CERK, while PI3K?CAkt promotes survival and proliferation. Upstream regulators like TGF-??, EGF, TNF-??, IL-1??, and mechanical strain modulate ITGA2 expression. Downstream, the pathway drives MYC and CCND1 transcription, MMP-mediated matrix remodeling, and focal adhesion reinforcement through talin, paxillin, vinculin, and kindlin.
In colorectal cancer, ITGA2-mediated ECM adhesion supports tumor cell survival in the microenvironment and facilitates basement membrane invasion. HT29 cells typically have wild-type APC and TP53, enabling study of ??-catenin?Cindependent adhesion. ITGA2 knockout in this context allows dissection of ??2??1 integrin contributions to anoikis resistance, collective migration, and collagen I signaling, independent of other integrins. This model is valuable for investigating integrin?CECM crosstalk in inflammatory bowel disease?Cassociated fibrosis, thrombosis due to altered platelet collagen receptors, and metastatic colonization.
Key applications include collagen I adhesion assays, Transwell migration/invasion assays, and Western blotting for pFAK Y397 and pERK. Flow cytometry confirms loss of surface integrin ??2, immunofluorescence visualizes vinculin/paxillin at focal adhesions, and co-immunoprecipitation assesses ITGA2?CITGB1 interaction. Additional assays include RT-qPCR for transcript ablation, anoikis apoptosis assays, and collagen-based drug sensitivity screening. For further details, please contact Ascent Research.
