The DLG5 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the widely used HT29 human colorectal adenocarcinoma cell line. This product provides a genetically heterogeneous pool of cells with targeted disruption of the DLG5 gene, making it well-suited for studying gene function in a mixed population context. As a polyclonal knockout, it avoids clonal artifacts and is ideal for early-stage target validation and functional screening assays.
HT29 is a human colorectal adenocarcinoma cell line originally established from a primary tumor. It displays epithelial morphology and is widely utilized as an in vitro model for intestinal epithelial biology, cancer research, and drug discovery. These cells form polarized monolayers and express key tight junction proteins, making them particularly suited for studying barrier function and cell polarity. Moreover, HT29 cells are known for their ability to differentiate into enterocyte-like cells under specific culture conditions, further enhancing their relevance for intestinal barrier studies.
DLG5 encodes a membrane-associated guanylate kinase (MAGUK) scaffold protein that localizes to tight junctions and adherens junctions. It plays a critical role in maintaining epithelial cell polarity and junctional integrity by linking adhesion complexes to the actin cytoskeleton. DLG5 interacts directly with junctional proteins such as occludin, ZO-1, and claudin-2, and also modulates signaling pathways including Wnt and Hippo. In the Wnt pathway, DLG5 influences ??-catenin stability and TCF/LEF-mediated transcription, while in the Hippo cascade it affects YAP/TAZ subcellular localization and activity. Upstream regulators include Wnt ligands and SRC kinase, and downstream targets comprise junctional components and transcriptional co-activators YAP and TAZ. By scaffolding these molecular interactions, DLG5 coordinates cytoskeletal organization with signal transduction.
Disruption of DLG5 in HT29 cells is expected to compromise tight junction assembly and cell polarity, leading to increased paracellular permeability and altered epithelial architecture. This knockout model provides a physiologically relevant background to dissect the interplay between cell adhesion and oncogenic signaling pathways. In the colorectal cancer context, DLG5 loss may dysregulate ??-catenin and YAP/TAZ activity, potentially impacting proliferation, differentiation, and tumorigenicity. Such phenotypic changes are highly informative for studying inflammatory bowel disease and colitis-associated cancer.
Researchers can utilize this polyclonal knockout cell population to investigate mechanisms governing tight junction dynamics, epithelial barrier function, and cross-talk between Wnt and Hippo pathways. Typical downstream assays include western blotting for junctional proteins (occludin, ZO-1), immunofluorescence staining of tight junction markers, transwell epithelial permeability measurements, RT-qPCR profiling of downstream targets, wound healing migration assays, and flow cytometric analysis of apoptosis or proliferation. Moreover, these cells serve as a platform for screening small-molecule modulators of junctional integrity or drug sensitivity in colorectal cancer. For further details or to discuss custom solutions, please contact Ascent Research.
