AFAP1 Knockout HT29 Polyclonal Cells are a polyclonal knockout cell population generated by CRISPR/Cas9-mediated disruption of the AFAP1 gene in the HT29 human colorectal adenocarcinoma cell line. This heterogeneous pool provides a robust loss-of-function model for investigating AFAP1-dependent processes without the biases of clonal selection. The product is suitable for advanced studies of actin dynamics, cell motility, and epithelial biology, offering a versatile tool for researchers examining cytoskeletal regulation and signaling in colorectal cancer contexts.
HT29 cells were originally derived from a primary colorectal adenocarcinoma of a 44-year-old female and have become a well-established intestinal epithelial model. These cells form polarized monolayers, express tight junction proteins, and are widely employed in research on barrier function, drug transport, and colorectal cancer biology. Their epithelial characteristics make them particularly valuable for dissecting pathways that govern cell adhesion, migration, and tissue integrity under both normal and pathogenic conditions.
AFAP1 encodes actin filament-associated protein 1, an adaptor that physically links Src family kinases to the actin cytoskeleton. It acts as a scaffold facilitating Src-mediated phosphorylation of substrates such as cortactin and p130Cas, thereby promoting focal adhesion disassembly and actin reorganization. Upstream regulators include integrin receptors, EGFR, and Src kinase, while interacting proteins include actin, PKC, and cortactin. AFAP1 functions downstream of integrin activation and FAK autophosphorylation; its subsequent phosphorylation modulates actin filament bundling and focal adhesion turnover, and it influences the expression of matrix metalloproteinases, linking Src/FAK signaling to invasive behavior.
In the HT29 cellular context, AFAP1 knockout disrupts cell migration and invasion and may impair epithelial barrier function, reflecting the protein??s central role in actin remodeling and focal adhesion dynamics. This model is highly relevant for exploring mechanisms of colorectal cancer metastasis, drug resistance, and epithelial-to-mesenchymal transition, as well as inflammatory bowel disease where barrier dysfunction is a critical factor. By removing AFAP1, researchers can delineate its specific contributions to Src-driven motility and integrin-mediated adhesion within a human colorectal adenocarcinoma background.
Researchers can apply AFAP1 Knockout HT29 Polyclonal Cells in a variety of functional assays, including wound healing, transwell migration/invasion, immunofluorescence staining for actin stress fibers, western blotting for phospho-FAK and phospho-Src, TEER measurement for barrier integrity, RT-qPCR for matrix metalloproteinases, and cell adhesion assays. These experiments enable detailed dissection of AFAP1-dependent signaling nodes and their impact on cell motility, barrier function, and focal adhesion turnover. The polyclonal population offers a representative loss-of-function model avoiding clonal artifacts. For further information or to request a quote, please contact Ascent Research.
