DLGAP4 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population that eliminates DLGAP4 expression in the human embryonic kidney HEK293T cell line. This loss-of-function model targets DLGAP4, a scaffold protein that links PSD-95 (DLG4) to SHANK family proteins at glutamatergic synapses. The polyclonal format provides a heterogeneous pool of edited cells with targeted DLGAP4 disruption, enabling functional studies without clonal isolation. The model is optimal for dissecting DLGAP4-dependent molecular interactions and signaling in a transfectable system suited for viral packaging, biochemical assays, and imaging.
HEK293T cells are human embryonic kidney cells expressing SV40 large T-antigen, which supports episomal plasmid replication and enhances transient protein expression and viral production. This host background enables reconstitution of synaptic protein complexes and is extensively used for studying postsynaptic density assembly. Although non-neuronal, HEK293T cells offer robust transfectability and a well-characterized biochemical environment for examining protein?Cprotein interactions, post-translational modifications, and signaling dynamics underlying synaptic scaffolding. In DLGAP4 knockout studies, HEK293T provides a clean system to assess DLGAP4 ablation without neuronal-specific variables.
DLGAP4 (SAPAP4) encodes a scaffold protein that directly binds the PDZ domain of PSD-95 and the C-terminus of SHANK proteins, forming the core postsynaptic density tripartite complex. DLGAP4 is regulated by neuronal activity and calcium influx through PKC-mediated phosphorylation, acting downstream to cluster PSD-95, SHANK3, Homer1, and cortactin. It anchors AMPA receptors (via GluA1) and NMDA receptors (via NR1), modulating receptor trafficking and synaptic strength. DLGAP4 knockout disrupts these assemblies, impairing membrane?Ccytoskeleton linkage. This model allows dissection of altered protein networks involving SHANK1/2/3, Homer1, and cortactin, and effects on receptor surface expression and scaffold dynamics.
In HEK293T cells, the DLGAP4 polyclonal knockout reveals non-neuronal scaffolding functions and complex organization capacity in a simplified environment. Although lacking synapses, these cells retain signaling and trafficking pathways that permit reconstitution of postsynaptic-like assemblies upon co-expression of binding partners. The model is valuable for identifying DLGAP4-dependent interactions via co-immunoprecipitation and for studying scaffold complex formation biochemistry. It also enables analysis of how DLGAP4 influences PSD-95 and SHANK3 stability and modification independent of synaptic context, isolating intrinsic scaffold properties.
Applications include co-immunoprecipitation and immunofluorescence to map DLGAP4 interactomes, Western blotting and RT-qPCR to assess expression changes in PSD-95, SHANK3, and Homer1, and reconstitution studies of AMPA/NMDA receptor trafficking. The polyclonal knockout cells are a robust platform for investigating signaling pathways dysregulated in schizophrenia, autism spectrum disorder, and intellectual disability. For further information or to request a quotation, please contact Ascent Research.
