The DLG1 Knockout HT29 Polyclonal Cells represent a CRISPR/Cas9-mediated gene-disrupted polyclonal population derived from the HT29 human colorectal adenocarcinoma cell line. This polyclonal knockout product provides a pooled loss-of-function model for investigating DLG1-dependent cellular processes without the selection of a single clonal isolate, thereby preserving heterogeneous genetic backgrounds that may better recapitulate tumor heterogeneity.
HT29 cells, originally isolated from a 44-year-old female with colorectal adenocarcinoma, are adherent, epithelial-like cells of intestinal crypt origin. They constitutively express markers characteristic of intestinal epithelial cells and are extensively utilized as an in vitro model for studying intestinal epithelium biology, colorectal cancer pathogenesis, and drug response. Their well-differentiated phenotype under standard culture conditions makes them particularly suitable for analyzing epithelial barrier function and apicobasal polarity.
DLG1 (Discs Large Homolog 1) encodes a membrane-associated guanylate kinase (MAGUK) scaffold protein that is critical for the assembly and maintenance of cell?Ccell junctions, including tight junctions and adherens junctions. At the molecular level, DLG1 interacts with and helps localize key proteins such as E-cadherin, ??-catenin, ZO-1, APC, and PTEN to the plasma membrane. It functions downstream of calcium influx and is regulated by upstream factors including APC, PTEN, protein kinase C, and Src family kinases. Through these interactions, DLG1 modulates Wnt/??-catenin signaling, Hippo pathway activity, and PI3K-Akt signaling, thereby coordinating cell polarity, proliferation, and migration. Notably, its association with APC and PTEN positions DLG1 as a pivotal node in the negative regulation of ??-catenin transcriptional activity.
In the context of HT29 colorectal adenocarcinoma cells, DLG1 knockout is expected to disrupt tight junction integrity and apical?Cbasal polarity, leading to altered epithelial barrier function and dysregulated Wnt signaling. Given that HT29 cells harbor an activating ??-catenin mutation, the additional loss of DLG1, which normally scaffolds the APC destruction complex, may further enhance ??-catenin-dependent transcription and promote a more invasive phenotype. Thus, this model offers a powerful tool to dissect how loss of cell polarity contributes to colorectal cancer progression, metastasis, and chemoresistance.
Researchers can employ this polyclonal knockout cell population in a wide range of functional assays, including quantitative RT-qPCR and western blotting to confirm DLG1 disruption, co-immunoprecipitation and immunofluorescence to study altered protein localization, transepithelial electrical resistance (TEER) measurements to assess barrier function, ??-catenin reporter assays to gauge Wnt pathway activity, and migration/invasion assays to explore metastatic potential. It is also suitable for RNA-seq-based transcriptomic profiling and drug screening campaigns aimed at identifying compounds that target polarity-deficient tumor cells. For additional technical details, please contact Ascent Research.
