GPR39 CRISPR Activation Plasmid (h): sc-410802-ACT

Human GPR39 encodes a zinc-sensing G protein–coupled receptor that integrates extracellular Zn2+ availability with intracellular signaling to modulate cellular excitability, secretion, and survival programs. Upon activation, GPR39 can couple to Gq/11- and Gs-associated pathways, promoting phospholipase C–dependent calcium signaling and cAMP/PKA cascades that intersect with MAPK/ERK and transcriptional responses. These signaling outputs have been linked to regulation of epithelial and metabolic homeostasis, inflammatory signaling, and neuronal function, making GPR39 a useful node for studying GPCR-driven pathway dynamics. Altered GPR39 expression or signaling has been associated in the literature with cardiometabolic phenotypes, gastrointestinal barrier processes, and neurobehavioral endpoints, supporting its relevance for mechanistic disease modeling in cell-based systems.

GPR39 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GPR39 expression without altering the underlying DNA sequence.

GPR39 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GPR39 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 GPR39 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GPR39 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GPR39 locus and enabling the study of GPR39-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GPR39 pathway restoration in tumor cells with silenced or reduced GPR39 expression.

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