The ACTA2 Knockout CAL-27 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the human tongue squamous cell carcinoma line CAL-27. This product provides a heterogeneous pool of cells with targeted disruptions in ACTA2, enabling functional studies of ??-smooth muscle actin (??-SMA) without clonal selection bias. As a polyclonal knockout model, it is suited for population-level assays and avoids artifacts from single-cell cloning.
CAL-27 is an adherent epithelial cell line from a human tongue squamous cell carcinoma, widely used in oral cancer research for its tumorigenic and invasive properties. This cell line exhibits strong responses to TGF-?? stimulation, which induces myofibroblastic differentiation and upregulates ACTA2. The knockout model in CAL-27 allows dissection of ??-SMA-dependent phenotypes in a relevant epithelial tumor microenvironment.
ACTA2 encodes ??-SMA, a cytoskeletal protein that polymerizes into contractile stress fibers and interacts with myosin, tropomyosin, vinculin, and other focal adhesion and contractile components. Its transcription is driven by TGF-??1 via SMAD2/3 and serum response factor (SRF) with co-activators MRTF-A/B, and is also sensitive to mechanical stress. Downstream, ??-SMA mediates cell contractility, migration, and extracellular matrix remodeling. Knockout of ACTA2 disrupts this central node, impairing TGF-??-induced stress fiber formation and matrix contraction.
In CAL-27 cells, TGF-??1 stimulation promotes a myofibroblast-like phenotype characterized by enhanced ??-SMA expression, collagen contraction, and invasiveness. The ACTA2 knockout polyclonal population abrogates this response, providing a powerful tool to separate ??-SMA-mediated effects from other TGF-?? pathways. This model is valuable for studying epithelial-to-mesenchymal transition (EMT) and tumor cell plasticity, as well as the role of tumor-derived ??-SMA in stromal remodeling, without confounding signals from fibroblast populations.
Applications include western blotting and immunofluorescence for ??-SMA and stress fibers, RT-qPCR, scratch-wound and transwell assays for migration and invasion, collagen gel contraction, and adhesion assays. The model is especially suited for TGF-?? stimulation experiments and RNA-seq transcriptomics to identify ACTA2-dependent gene programs. It supports anti-fibrotic drug screening, cancer-associated fibroblast activation studies, and tumor microenvironment research. For further details, please contact Ascent Research.
