GALR2 CRISPR Activation Plasmid (h): sc-403779-ACT

Human GALR2 encodes galanin receptor 2 (GALR2), a class A GPCR that binds the neuropeptide galanin to modulate neuronal excitability, neurotransmitter release, and neuroendocrine secretion. Upon ligand engagement, GALR2 primarily couples to Gq/11 to activate PLCβ, IP3/DAG signaling, intracellular Ca2+ mobilization, and PKC-dependent pathways, with additional context-dependent coupling to other G proteins that shapes MAPK/ERK signaling and gene expression programs. These signaling outputs link GALR2 to regulation of pain processing, mood-related behaviors, feeding and energy balance, and smooth muscle contractility across peripheral tissues. Dysregulated GALR2 expression or signaling has been investigated in neuropsychiatric and metabolic phenotypes and in tumor biology where GPCR-driven pathways can influence proliferation, survival, and migration, supporting its value as a pathway probe in mechanistic studies.

GALR2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GALR2 expression without altering the underlying DNA sequence.

GALR2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GALR2 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 GALR2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GALR2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GALR2 locus and enabling the study of GALR2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GALR2 pathway restoration in tumor cells with silenced or reduced GALR2 expression.

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