The DLG1 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HEK293T cell line. These cells feature targeted disruption of the DLG1 gene, which encodes the discs large homolog 1 scaffold protein. This polyclonal pool provides a heterogeneous loss-of-function model suitable for investigating DLG1-dependent cellular processes without the limitations of single-cell clonal selection.
HEK293T cells are immortalized human embryonic kidney epithelial cells that constitutively express the SV40 large T-antigen, enabling episomal replication of plasmids containing the SV40 origin of replication. Originally derived from the HEK293 parental line, HEK293T is a widely used host for transient and stable transfections, lentiviral packaging, and protein production due to its high transfectability and robust expression capacity. The cells exhibit adherent growth and rapid proliferation, facilitating scalable experimental workflows.
DLG1 functions as a multidomain scaffold protein of the membrane-associated guanylate kinase (MAGUK) family, orchestrating the assembly of macromolecular signaling complexes at cell?Ccell junctions and synapses. Through its PDZ, SH3, and GUK domains, DLG1 interacts with key partners including CASK, LIN7, and MPP2, and is regulated by upstream kinases such as CaMKII and Src. DLG1 is integral to Wnt and Hippo signaling: it scaffolds Axin, APC, and GSK3B to modulate ??-catenin/TCF/LEF-mediated transcription, and influences YAP/TAZ activity through tight junction?Cassociated complexes. DLG1 also controls AMPA receptor trafficking by binding GRIA1 and GRIA2 subunits, and stabilizes E-cadherin/??-catenin adhesion complexes while regulating PTEN localization and p38 MAPK activation.
Although HEK293T cells do not form fully polarized epithelial monolayers, they express a repertoire of junctional and signaling proteins, making them a convenient system for dissecting DLG1 functions independent of mature epithelial architecture. DLG1 disruption in this background enables focused analysis of Wnt and Hippo pathway transduction, AMPA receptor surface expression, and cytoskeletal rearrangement. The polyclonal knockout format captures a spectrum of genetic perturbations, allowing researchers to assess phenotypic variability and identify robust DLG1-dependent mechanisms.
These cells are suitable for a wide range of experimental approaches, including Western blotting and immunofluorescence to confirm protein loss and localization, co-immunoprecipitation and RT-qPCR to assess protein interactions and transcript levels. Further functional assays include TOPFlash or TEAD reporter assays to measure Wnt and Hippo pathway activity, cell migration and proliferation assays to model tumor suppression, and flow cytometry for surface receptor analysis. The model supports investigations into neurodevelopmental disorders, inflammatory bowel disease, and cancer metastasis. For further details, please contact Ascent Research.
