GRPR CRISPR Activation Plasmid (h): sc-401637-ACT

Human GRPR encodes the gastrin-releasing peptide receptor, a seven-transmembrane GPCR that binds gastrin-releasing peptide and related bombesin-like ligands to regulate neuroendocrine signaling. GRPR activation engages heterotrimeric G proteins to stimulate PLCβ-dependent phosphoinositide turnover, intracellular Ca2+ mobilization, and downstream MAPK/ERK and PKC signaling, influencing proliferation, secretion, and smooth muscle contractility. The receptor is broadly relevant to studies of sensory processing and gastrointestinal physiology, and altered GRPR signaling has been linked to oncogenic and neurobehavioral phenotypes in multiple model systems. GRPR expression and pathway activity are frequently used to interrogate GPCR-driven transcriptional programs, calcium dynamics, and ligand-dependent signaling biases in human cells.

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

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

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