G_γ13 CRISPR Activation Plasmid (h): sc-403045-ACT

GNG13 encodes the human G protein gamma subunit Gγ13, a component of heterotrimeric G protein complexes that couple activated GPCRs to downstream effector pathways. As part of the Gβγ dimer, Gγ13 helps regulate signal propagation to targets such as phospholipase Cβ and ion channels, shaping second-messenger outputs including calcium flux and MAPK/ERK responses. GNG13 expression is enriched in sensory-associated contexts, supporting roles in chemosensory and neuronal signaling where precise GPCR coupling is required. Dysregulated G protein subunit expression or GPCR pathway remodeling is frequently observed in cancer and neurobiology-relevant states, making GNG13 useful for mechanistic studies of signaling plasticity and cell-state control.

Gγ13 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GNG13 expression without altering the underlying DNA sequence.

Gγ13 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GNG13 locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.

Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the GNG13 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Gγ13 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GNG13 locus and enabling the study of Gγ13-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Gγ13 pathway restoration in tumor cells with silenced or reduced GNG13 expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.