The DLG1 Knockout HCT 116 Polyclonal Cells comprise a mixed population of the HCT 116 colorectal carcinoma cell line engineered via CRISPR/Cas9 to disrupt the DLG1 gene. This polyclonal format minimizes clonal artifacts and provides a biologically diverse knockout model suitable for studying DLG1 loss-of-function. The pool lacks wild-type DLG1 expression, enabling investigation of cellular consequences without single-cell selection.
HCT 116 is an epithelial line from human colorectal carcinoma, harboring a KRAS G13D oncogenic mutation and a stabilizing CTNNB1 (??-catenin) mutation. The latter drives constitutive TCF/LEF-dependent transcription, creating a Wnt-activated background. Combined with KRAS-driven MAPK signaling, these cells represent a well-characterized model for colorectal tumorigenesis, particularly suited to analyzing factors that modulate Wnt pathway output and epithelial integrity.
DLG1 functions as a scaffold protein at tight and adherens junctions, recruiting factors such as APC, PTEN, LIN7, CASK, and ERBIN to regulate polarity and suppress proliferation. It promotes ??-catenin degradation by facilitating APC complex assembly, thereby restraining Wnt signaling. DLG1 is phosphorylated by CDK5 and SRC, which control its junctional localization. Its loss disrupts APC/??-catenin interaction, enhances TCF/LEF transcriptional activity, and leads to mislocalization of PTEN and tight junction components TJP1, CLDN, and OCLN, while also affecting Hippo pathway effectors YAP/TAZ.
In the HCT 116 background, the stabilizing CTNNB1 mutation sensitizes cells to DLG1-mediated Wnt regulation; knockout therefore exaggerates Wnt target gene expression and disrupts cell polarity programmes. This synergy between oncogenic KRAS, mutant ??-catenin, and DLG1 loss mirrors aggressive colorectal cancer phenotypes. The model enables dissection of how polarity defects and hyperactive Wnt/??-catenin signaling collaborate to drive proliferation, migration, and loss of barrier function.
Typical applications include western blotting and RT-qPCR for DLG1 and target molecule validation, co-immunoprecipitation for complex analysis, immunofluorescence for protein localization, proliferation and migration assays, TOP/FOPflash reporters for Wnt activity, and TEER for tight junction integrity. These tools support research into tumor suppressor mechanisms, cell adhesion, Wnt pathway inhibition, and junction biology. For additional details or custom cell solutions, contact Ascent Research.
