The EHBP1 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population generated from the human HT29 colorectal adenocarcinoma cell line. This product provides a loss-of-function model through targeted disruption of the EHBP1 (EH domain-binding protein 1) gene, enabling researchers to investigate the functional consequences of EHBP1 deficiency in intestinal epithelial cells.
HT29 cells are a well-characterized human colorectal adenocarcinoma cell line with epithelial morphology. These cells serve as a robust model for studying intestinal epithelial barrier function, drug absorption mechanisms, and colorectal cancer biology. Their ability to form polarized monolayers and express enterocytic markers makes them particularly valuable for investigating epithelial cell signaling, membrane trafficking, and metabolic regulation.
EHBP1 functions as an adapter protein that links clathrin-mediated endocytosis to actin cytoskeleton dynamics. It is activated downstream of insulin signaling through the PI3K/AKT pathway and interacts with EHD1 and EHD2 to coordinate endocytic vesicle trafficking. EHBP1 directly binds to actin and clathrin, facilitating complex formation with Rab family GTPases to regulate membrane remodeling. A critical function of EHBP1 is its role in insulin-stimulated GLUT4 translocation to the plasma membrane, where it promotes glucose uptake. The mechanistic pathway involves insulin receptor activation, PI3K-dependent AKT phosphorylation, and recruitment of EHD1/EHBP1 complexes that orchestrate actin polymerization and vesicle fusion, ultimately driving GLUT4 insertion. Disruption of EHBP1 impairs this process, leading to defective glucose transport and altered membrane trafficking.
In HT29 colorectal adenocarcinoma cells, EHBP1 disruption provides a physiologically relevant system to dissect the interplay between insulin signaling, endocytic trafficking, and epithelial cell function. HT29 cells express key components of the insulin pathway, including the insulin receptor, PI3K, and AKT, and exhibit insulin-regulated glucose transport, making them suitable for studying EHBP1-dependent GLUT4 translocation. Moreover, EHBP1??s role in actin dynamics and membrane trafficking is critical for maintaining epithelial polarity and cell migration, processes often dysregulated in colon cancer progression. Thus, this knockout model enables investigation of EHBP1??s contribution to both metabolic and oncogenic phenotypes within an intestinal epithelial context.
Researchers can employ this polyclonal knockout population in a variety of experimental settings, including insulin-stimulated glucose uptake assays, GLUT4 translocation analysis by immunofluorescence, and transferrin uptake assays to assess clathrin-mediated endocytosis. The cells are also suitable for actin cytoskeleton staining to evaluate EHBP1-dependent cytoskeletal rearrangements and cell migration assays relevant to cancer metastasis. Additionally, this model supports drug target screening aimed at restoring insulin sensitivity or inhibiting aberrant endocytosis in colorectal cancer. For further information, please contact Ascent Research.
