The GNAZ Knockout HEK293T Polyclonal Cells are a heterogeneous population of HEK293T cells with CRISPR/Cas9-mediated disruption of the GNAZ gene, providing a loss-of-function model for the inhibitory G??z subunit. This polyclonal knockout pool retains the host cell line??s genetic background while introducing targeted gene disruption across the population, enabling physiologically relevant interrogation of G??z-dependent signaling without clonal biases. The polyclonal format minimizes clonal artifacts and preserves population-level signaling dynamics, facilitating studies of compensatory mechanisms and network-level responses.
The HEK293T host cell line is a human embryonic kidney epithelial line transformed with adenovirus type 5 DNA and expressing the SV40 large T antigen. These modifications impart high proliferative capacity, exceptional transfectability, and robust protein expression, establishing HEK293T as a standard platform for GPCR signaling, functional genomics, and protein interaction studies. Its epithelial phenotype and complemented antigen expression support diverse experimental protocols, and the cells endogenously express many signaling components relevant to G protein cascades.
GNAZ encodes G??z, a pertussis toxin-insensitive Gi/o family G?? subunit that inhibits adenylyl cyclase, reducing cAMP levels. It is activated by Gi/o-coupled receptors such as alpha-2 adrenergic, D2 dopamine, 5-HT1A serotonin, and melatonin receptors. Downstream targets include adenylyl cyclase, cAMP, PKA, CREB, and MAPK cascades. G??z interacts with G?¦? subunits, RGS proteins (RGS4, RGS19), and GPCR kinases. The pathway axis GPCR ?? G??z ?? adenylyl cyclase ?? cAMP ?? PKA ?? CREB underscores its role in negative cAMP regulation and cross-talk with MAPK/ERK and PI3K-Akt signaling.
Knockout of GNAZ in HEK293T removes G??z-mediated adenylyl cyclase inhibition, altering basal and agonist-stimulated cAMP production. This model dissects G??z-specific contributions from overlapping Gi/o signaling, allowing discrimination of G??z-dependent outputs and investigation of compensatory RGS or G protein adaptations. It offers a human cellular context to examine native GPCR dynamics without exogenous receptor overexpression, supporting studies of biased signaling and pathway crosstalk.
Applications include mechanistic GPCR signaling studies, cAMP modulation screens, pharmacological profiling of Gz-coupled receptors, and functional genomics mapping of G??z-dependent transcriptional networks. Validated assays comprise Western blot for G??z, cAMP ELISA, RT-qPCR and RNA-seq for transcriptional changes, CREB phosphorylation analysis, CRE-luciferase reporter assays, co-immunoprecipitation, and flow cytometry. These cells serve drug discovery targeting Gz-mediated pathways. For technical details, contact Ascent Research.
