The GNA13 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the GNA13 gene in a HEK293T background. This loss-of-function model eliminates endogenous G??13 expression, enabling study of its specific signaling roles. The polyclonal format maintains genetic heterogeneity while achieving effective gene disruption across the population, making it suitable for pooled functional assays and reducing artifacts from single-cell selection.
HEK293T cells, derived from HEK293 by stable integration of the SV40 large T antigen, are a widely used human embryonic kidney epithelial line with adherent morphology. They offer high transfection efficiency, robust protein overexpression, and are preferred for lentiviral and retroviral production. The parental line expresses numerous endogenous GPCRs, providing a physiologically relevant context for dissecting G??13-dependent signaling downstream of diverse stimuli.
GNA13 encodes G??13, a mediator of GPCR signals to RhoA. Ligand engagement of receptors such as S1PR1?C5, PAR1, and LPAR1?C3 triggers GTP loading on G??13, causing its dissociation from G?¦? and subsequent activation of RhoGEFs including p115-RhoGEF (ARHGEF1), LARG (ARHGEF12), and PDZ-RhoGEF (ARHGEF11). These GEFs stimulate RhoA, which activates ROCK1/2 to phosphorylate MLC2 and LIMK, leading to cofilin inactivation and actin polymerization. This cascade promotes stress fiber formation, focal adhesion assembly, and cell migration. Parallel RhoA signaling via SRF and MRTF-A drives transcriptional responses. Scaffolds like spinophilin and RGS domain-containing RhoGEFs fine-tune the signaling complex.
In HEK293T cells, GNA13 knockout disrupts a pivotal node connecting GPCR inputs to RhoA-driven cytoskeletal and transcriptional outputs. This model enables dissection of G??13-specific functions, separating them from G??q and G??12 pathways. Given its roles in platelet activation, cardiac hypertrophy, sensorineural hearing loss, and cancer metastasis??including diffuse large B-cell lymphoma and solid tumor invasion??the knockout cells offer a versatile platform for mechanistic studies. The HEK293T background facilitates high-efficiency transfection for reconstitution experiments and reporter assays, allowing precise validation of G??13 interactors and downstream effectors.
GNA13 knockout HEK293T polyclonal cells support a range of assays: GTP-RhoA pull-down to measure RhoA activation, phospho-MLC2 and phospho-cofilin immunoblotting, SRE-luciferase reporter assays for SRF activity, wound-healing and transwell migration/invasion assays, and phalloidin staining for actin cytoskeletal imaging. Co-immunoprecipitation confirms interactions between G??13 and RhoGEFs, while RT-qPCR quantifies SRF target genes. The model also enables high-throughput screening of GPCR modulators targeting G??13-dependent pathways. For further technical information, please contact Ascent Research.
