The DLGAP4 Knockout HeLa Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population originating from the widely utilized HeLa cell line, with targeted disruption of the DLGAP4 gene. This gene-edited product provides a mixed population of cells carrying loss-of-function mutations in DLGAP4, enabling robust functional studies without the clonal selection bottlenecks inherent to single-cell-derived knockout lines. The polyclonal format preserves phenotypic heterogeneity, making it particularly suitable for experiments that require a representation of the genetic diversity observed in unselected populations, such as drug response assays or pooled functional screens.
The HeLa cell line, established from a cervical adenocarcinoma in 1951, is an immortalized epithelial cell model extensively employed in cancer biology, virology, cell signaling, and toxicology. These cells are characterized by their robust growth kinetics, ease of culture, and a well-annotated genomic landscape. As a cancer-derived line, HeLa cells exhibit aberrant signaling pathways, including those governing cell adhesion, cytoskeletal dynamics, and proliferation, making them an ideal host for interrogating the tumor-suppressive or oncogenic functions of genes like DLGAP4.
DLGAP4 encodes a postsynaptic density (PSD) scaffold protein that bridges membrane-associated guanylate kinases, such as DLG4/PSD95, to the actin cytoskeleton via interactions with SHANK family proteins. It plays a critical role in the assembly and maintenance of macromolecular signaling complexes at cell junctions, coordinating glutamatergic signaling, postsynaptic organization, and actin remodeling. In the context of the HeLa cell model, DLGAP4 is implicated in epithelial cell adhesion and migration through its interactions with cadherin complexes and focal adhesion kinase (FAK). Upstream regulators include SRC family kinases, Ca2+/calmodulin-dependent kinase II (CAMK2), and protein kinase C, while downstream targets encompass DLG4, SHANK1, NMDA receptor subunits (GRIN1, GRIN2B), and effectors of actin polymerization. The DLGAP4 scaffold integrates signals from cell surface receptors to cytoskeletal reorganization, thereby influencing cellular morphology and motility.
In HeLa cells, DLGAP4 knockout is anticipated to perturb the linkage between membrane proteins and the actin cytoskeleton, leading to compromised cell adhesion, altered spreading, and enhanced migration??phenotypes associated with metastatic cancer cells. This model enables the dissection of DLGAP4-dependent pathways that intersect with oncogenic signaling networks, providing a platform to study how scaffold protein dysfunction contributes to epithelial-to-mesenchymal transition and tumor progression. The polyclonal nature of the knockout population allows for the assessment of heterogeneous responses to pathway perturbations, which is especially relevant for understanding clonal evolution in cancer.
This DLGAP4 knockout product is well-suited for a broad spectrum of research applications, including functional characterization of scaffold proteins in cancer biology, interactomic mapping of the DLGAP4-DLG4-SHANK1 complex, and high-content screening for compounds that modulate cell adhesion or migration. Routine characterization can be performed using western blotting, RT-qPCR, and immunofluorescence, while functional interrogation can be achieved through cell migration assays, proliferation assays, co-immunoprecipitation, and phospho-signaling analyses. For detailed technical specifications or to discuss customization options, please contact Ascent Research.
