GPR19 CRISPR Activation Plasmid (h): sc-407000-ACT

GPR19 encodes an orphan G protein-coupled receptor (GPCR) that is expressed in multiple tissues and has been linked to regulation of cellular signaling networks that shape transcriptional programs, differentiation state, and stress-responsive physiology. As a membrane receptor, GPR19 is positioned to influence GPCR-coupled pathways such as cAMP/PKA and MAPK signaling, with downstream effects on gene expression and cellular homeostasis. Altered GPCR signaling has broad relevance to metabolic regulation, neurobiology, and immune modulation, and GPR19 expression changes have been investigated in contexts including inflammation and cancer-associated phenotypes. Modulating endogenous GPR19 levels supports mechanistic studies of receptor-driven signaling dynamics and pathway cross-talk in human cell models.

GPR19 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GPR19 expression without altering the underlying DNA sequence.

GPR19 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GPR19 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 GPR19 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GPR19 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GPR19 locus and enabling the study of GPR19-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GPR19 pathway restoration in tumor cells with silenced or reduced GPR19 expression.

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