The DLG3 Knockout HAP1 Polyclonal Cells product is a polyclonal population of HAP1 cells with CRISPR/Cas9-mediated disruption of the DLG3 gene, which encodes the MAGUK scaffold protein SAP102. This targeted gene disruption creates a loss-of-function model for studying postsynaptic organization. The polyclonal format ensures a heterogeneous mix of edited alleles, supporting robust functional assessments without clonal selection. This tool is designed for researchers exploring synaptic scaffolding mechanisms and neurodevelopmental disorders.
The host HAP1 cell line is a human near-haploid hematopoietic line derived from the KBM-7 chronic myeloid leukemia (CML) cell line. Its near-haploid karyotype simplifies gene editing, requiring disruption of a single allele, and the cells grow in suspension, facilitating high-throughput genetic screening. Although non-neuronal, HAP1 cells express synaptic scaffold proteins, making them suitable for studying DLG3 interactions in a reductionist system.
DLG3 encodes SAP102, a member of the membrane-associated guanylate kinase (MAGUK) family that clusters NMDA receptors (GRIN2A, GRIN2B) and AMPA receptors (GRIA1) at excitatory synapses. SAP102 also anchors potassium channels (KCNA4) and bridges to downstream signaling molecules via interactions with other MAGUKs such as DLG4 and DLG1, cell adhesion molecules like neuroligin (NLGN1), and scaffold adaptors including DLGAP1 and the SHANK family. Its scaffolding activity is dynamically regulated by neuronal activity, calcium influx, and phosphorylation by CaMKII and PKA, which modulate its binding affinity and receptor retention at the postsynaptic density.
In the HAP1 cellular environment, DLG3 knockout provides a simplified biochemical model for studying SAP102 function, circumventing the complexity of intact neuronal synapses. HAP1 cells endogenously express glutamate receptor subunits and key MAGUK partners, enabling detailed investigation of protein?Cprotein interactions, turnover, and post-translational modifications in a genetically tractable and high-throughput-compatible system. This model is particularly useful for testing how SAP102 loss destabilizes receptor clustering and alters glutamatergic signaling components, molecular events that contribute to X-linked intellectual disability, autism spectrum disorders, and schizophrenia.
The DLG3 Knockout HAP1 Polyclonal Cells support diverse assays: western blotting to quantify SAP102 and synaptic proteins, co-immunoprecipitation to profile protein complex composition, and immunofluorescence to visualize scaffold localization. RT-qPCR can measure transcriptional changes in downstream genes like GRIN2A and GRIA1, while calcium imaging assesses functional signaling deficits. This model is suitable for drug screening targeting cognitive impairment, synaptic plasticity research, and mechanistic dissection of neurodevelopmental disease pathways. For additional information, please contact Ascent Research.
