The DLGAP4 Knockout A-549 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the DLGAP4 gene within the A-549 human lung adenocarcinoma cell line. This heterogeneous pool of edited cells serves as a loss-of-function model, enabling functional studies without the requirement for clonal isolation. The polyclonal format provides a cost-effective and robust platform for assessing pooled effects of target gene disruption, making it suitable for high-throughput screening and pathway analysis in a cancer-relevant cellular context.
The A-549 host cell line, originally derived from a 58-year-old Caucasian male with lung adenocarcinoma, is a widely used model for non-small cell lung cancer research. These cells exhibit hypodiploid epithelial morphology and possess well-characterized genomic features that facilitate targeted gene editing and phenotypic characterization. A-549 cells are routinely employed in studies of tumor biology, metastasis, and drug sensitivity, offering a reliable and reproducible system for CRISPR-based applications.
DLGAP4 encodes a key scaffolding protein of the SAPAP/GKAP family that organizes the postsynaptic density at excitatory synapses. Mechanistically, DLGAP4 functions as a central adaptor, binding directly to PSD-95 and linking it to the SHANK1/2/3 and Homer protein complexes to stabilize clusters of ionotropic glutamate receptors, including GluN1 and GluA1 subunits. This assembly is critical for efficient glutamatergic signaling and synaptic plasticity. DLGAP4 is regulated by synaptic activity, with NMDA receptor activation, CaMKII, and PKA modulating its function, and it interacts with neuroligins to influence actin cytoskeleton remodeling and cell adhesion.
Although DLGAP4 is predominantly studied in neural tissues, its expression in non-neuronal contexts suggests broader roles in cell signaling. In A-549 adenocarcinoma cells, loss of DLGAP4 may disrupt cell junction organization and adhesion-dependent pathways, potentially impacting proliferation, migration, or tumor suppression. This model allows researchers to explore the functional repurposing of synaptic scaffold proteins in cancer, addressing whether DLGAP4 contributes to oncogenic processes or represents a vulnerability in lung cancer cells.
Researchers can utilize these polyclonal knockout cells in diverse assays: western blotting with anti-DLGAP4 antibodies and RT-qPCR verify protein and mRNA ablation; Sanger sequencing confirms target locus disruption; immunocytochemistry reveals altered subcellular distribution; and co-immunoprecipitation assays assess PSD-95 complex integrity. Functional studies include MTT proliferation, Transwell migration/invasion, and RNA-sequencing for transcriptome-wide effects. These cells are also ideal for high-content screening to identify modulators of scaffold interactions. For further details, please contact Ascent Research.
