This product is a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HCT 116 human colorectal carcinoma line, with targeted disruption of the DLG3 gene. DLG3, encoding a membrane-associated guanylate kinase (MAGUK) scaffold protein, is mutated in a pooled format to create a loss-of-function model without specification of individual editing events. The heterogeneous population is suitable for investigating DLG3-dependent processes in cell signaling, adhesion, and cancer biology.
The HCT 116 host cells are established from human colorectal carcinoma and carry an activating KRAS G13D mutation alongside a stabilizing CTNNB1 (??-catenin) mutation, resulting in constitutive Wnt pathway activity. They exhibit microsatellite stability (MSS) and wild-type TP53. These characteristics provide a robust genetic background for studying the intersection of oncogenic signaling and scaffold protein function, particularly in Wnt-driven colorectal carcinogenesis and preclinical drug response evaluation.
DLG3 functions as a molecular scaffold organizing multi-protein complexes at cell?Ccell junctions and synapses. It interacts with APC, ??-catenin, NMDA receptor subunits (GRIN2A, GRIN2B), and potassium channels (KCNA4). Upstream regulators include Wnt ligands (Wnt3a), SRC kinases, and protein kinase C; downstream targets are ??-catenin, TCF/LEF factors, and ionotropic receptors. DLG3 scaffolds the ??-catenin destruction complex and modulates TCF/LEF transcriptional output, integrating cell adhesion and Wnt signaling.
In the HCT 116 background, mutationally activated Wnt/??-catenin signaling is rendered independent of ligand, yet scaffolding by DLG3 may still influence pathway output by sequestering ??-catenin or altering its nuclear translocation. Disruption of DLG3 can therefore dissect the contribution of junctional scaffolding to tumor cell behavior, including proliferation, migration, and cell?Ccell adhesion, and help evaluate its proposed tumor-suppressor role in colorectal cancer.
Research applications feature Wnt luciferase reporter assays (TOPFlash/FOPFlash), co-immunoprecipitation with APC and ??-catenin, immunofluorescence for ??-catenin localization, and functional assays such as MTT proliferation, colony formation, and wound-healing/transwell migration. This polyclonal knockout pool is also amenable to high-throughput functional genomics screening and drug discovery campaigns targeting Wnt-addicted colorectal cancers. For additional information, please contact Ascent Research.
