The GPRIN3 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population derived from human embryonic kidney HEK293T cells, featuring disruption of the GPRIN3 gene. This product provides a genetically heterogeneous knockout model designed for loss-of-function studies of GPRIN3, a G protein-regulated inducer of neurite outgrowth. By eliminating GPRIN3 expression, the cells facilitate dissection of signaling pathways downstream of G protein-coupled receptors (GPCRs). The polyclonal format avoids the clonal selection bottleneck, offering a more representative population for functional genomics and signal transduction research.
The host cell line, HEK293T, is an adherent embryonic kidney epithelial line immortalized with the SV40 large T antigen, enabling high-level transient protein expression and efficient viral production. It supports stable transfection and genome editing applications, making it a workhorse for molecular and cellular biology. HEK293T cells endogenously express many GPCR signaling components, providing a biochemically accessible background to study cytoskeletal regulation and neurite outgrowth mechanisms.
GPRIN3 acts as a downstream effector of GPCR-activated G proteins, specifically interacting with G?? subunits GNAO1, GNAI1, and GNAZ. Upon receptor stimulation, GPRIN3 triggers neurite extension and actin cytoskeleton remodeling by activating the Rho GTPases RAC1 and CDC42. This stimulates a cascade involving PAK and LIMK kinases, which phosphorylate cofilin to promote actin filament dynamics. GPRIN3 also binds tubulin and actin, linking membrane-proximal events directly to cytoskeletal reorganization. The canonical pathway proceeds through GPCR??G????GPRIN3??RAC1??PAK1??LIMK??cofilin.
In HEK293T cells, GPRIN3 knockout eliminates this signaling hub, enabling precise analysis of how GPCRs and their cognate G proteins modulate actin-based processes. The polyclonal knockout population is particularly valuable for contrastive studies with wild-type controls, as it preserves natural cell-to-cell variability and reduces artifacts from single-cell cloning. This model supports investigation of G protein-coupling specificity and the interplay between GPRIN3 and other actin regulators in a non-neuronal context, where baseline neurite outgrowth machinery is absent unless introduced.
Research applications include GPCR signaling interrogation using luciferase reporters and cAMP assays, cytoskeletal visualization by phalloidin-based actin staining, and protein interaction mapping through co-immunoprecipitation and western blotting. RT-qPCR and immunofluorescence can monitor downstream gene expression and protein localization changes. The cells are also suited for heterologous expression of neuronal factors to reconstitute neurite outgrowth pathways in a controlled manner. For additional information, please contact Ascent Research.
