G_{β 2} CRISPR Activation Plasmid (h): sc-402936-ACT

GNB2 encodes the human G protein beta 2 (Gβ2) subunit, a core component of heterotrimeric G proteins that couple activated G protein–coupled receptors (GPCRs) to intracellular effectors. By partnering with Gγ subunits, Gβ2 helps regulate signaling outputs such as adenylyl cyclase activity, phospholipase Cβ activation, ion channel modulation, and downstream MAPK and PI3K-associated pathways that shape cell proliferation, migration, and neurotransmission. Altered GPCR/G protein signaling involving GNB2 has been associated with dysregulated signal transduction in neurological and oncogenic contexts, making it relevant for mechanistic studies of receptor-proximal signaling networks. As a widely expressed signaling scaffold, Gβ2 is often used to interrogate pathway crosstalk and stimulus-dependent transcriptional programs.

Gβ 2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GNB2 expression without altering the underlying DNA sequence.

Gβ 2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GNB2 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 GNB2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Gβ 2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GNB2 locus and enabling the study of Gβ 2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Gβ 2 pathway restoration in tumor cells with silenced or reduced GNB2 expression.

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