The GNA11 knockout HEK293T polyclonal cells comprise a CRISPR/Cas9-edited cell population in which the GNA11 gene has been disrupted to generate a functional knockout model of the G??11 protein. As a polyclonal pool, these cells harbor heterogeneous gene-editing events that collectively abrogate G??11 expression, making the population suitable for loss-of-function screens and pathway analysis without the need for single-cell cloning. This product provides a robust and cost-effective system for interrogating G??11-dependent signaling in an easily cultured and highly transfectable host background.
The parental HEK293T cell line is a human embryonic kidney-derived model transformed with adenovirus 5 DNA, which constitutively expresses the SV40 large T antigen, enabling episomal replication of transfected plasmids and high-level protein expression. Widely employed in protein production, viral packaging, and cell signaling research, HEK293T cells endogenously express many G protein-coupled receptors (GPCRs) and downstream effectors, making them a pertinent platform for studying G??11-mediated signal transduction. Their robust growth characteristics and amenability to transient and stable gene manipulation support downstream functional assays.
GNA11 encodes the G??11 subunit of heterotrimeric G proteins, a primary transducer of signals from Gq/11-coupled GPCRs. Upon receptor activation by ligands such as angiotensin II (AT1 receptor), endothelin-1 (ETA/ETB receptors), thrombin (PAR1), histamine (H1 receptor), and acetylcholine (muscarinic M1/M3/M5 receptors), G??11 exchanges GDP for GTP and activates phospholipase C ?? (PLC??). PLC?? hydrolyzes phosphatidylinositol 4,5-bisphosphate to generate inositol trisphosphate (IP3) and diacylglycerol (DAG). IP3 triggers calcium release from the endoplasmic reticulum, while DAG activates protein kinase C (PKC). Downstream, calcium/calmodulin-dependent kinase II (CaMKII) and the MAP kinase cascade (ERK1/2) propagate signaling. G??11 signaling is modulated by regulators of G protein signaling (RGS) proteins such as RGS2 and RGS4, and by interaction with G?¦? subunits and calmodulin.
In the HEK293T background, GNA11 knockout disrupts a central node of Gq/11-coupled calcium and ERK signaling, enabling researchers to dissect G??11-specific contributions versus those mediated by other G?? subunits or alternative pathways. This model is particularly relevant for studying oncogenic mutations in GNA11 that are prevalent in uveal melanoma and congenital vascular disorders such as Sturge-Weber syndrome and phakomatosis pigmentovascularis. The knockout cells allow for structure-function studies, mutant rescue experiments by reintroducing disease-associated variants, and screening of small-molecule inhibitors that target the G??11?CPLC?? axis. HEK293T??s high transfection efficiency facilitates such manipulations.
Typical applications of GNA11 knockout HEK293T polyclonal cells include quantitative measurements of GPCR-stimulated calcium flux using Fluo?4 AM, monitoring IP3 accumulation, PLC?? enzymatic activity assays, and western blot analysis of phospho-ERK1/2 and total ERK1/2. The cells are also suited for cell proliferation, migration, and invasion assays in cancer biology frameworks, as well as for identifying and validating pharmacological modulators of Gq/11 signaling. By serving as a G??11-null comparator, this model accelerates functional genomics pipelines and drug target discovery. For further information, protocols, or assistance with experimental design, please contact Ascent Research.
