GABA_B R2 CRISPR Activation Plasmid (h): sc-402517-ACT

GABBR2 encodes the GABA\(_B\) receptor subunit 2 (GABA\(_B\) R2), an essential component of functional metabotropic GABA\(_B\) receptors that form heterodimers with GABBR1 to mediate slow inhibitory neurotransmission in the central nervous system. Upon ligand binding, these receptors couple to Gi/o proteins to inhibit adenylyl cyclase, reduce cAMP/PKA signaling, modulate MAPK pathways, open GIRK potassium channels, and suppress voltage-gated calcium channel activity, collectively shaping neuronal excitability and synaptic plasticity. GABA\(_B\) signaling influences network oscillations, neurotransmitter release, and neuronal development, linking receptor regulation to multiple neurobiological processes. Altered expression or function of GABBR2 has been associated with neurological and neuropsychiatric phenotypes, including seizure susceptibility and dysregulated excitatory–inhibitory balance, making it relevant for mechanistic studies in disease models.

GABAB R2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GABBR2 expression without altering the underlying DNA sequence.

GABAB R2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GABBR2 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 GABBR2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GABAB R2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GABBR2 locus and enabling the study of GABAB R2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GABAB R2 pathway restoration in tumor cells with silenced or reduced GABBR2 expression.

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