GnRHR CRISPR Activation Plasmid (h): sc-401783-ACT

GNRHR encodes the human gonadotropin-releasing hormone receptor (GnRHR), a class A GPCR that mediates pituitary responses to hypothalamic GnRH to regulate luteinizing hormone and follicle-stimulating hormone secretion. Upon ligand binding, GnRHR primarily couples to Gq/11 to activate phospholipase C signaling, driving IP3/DAG production, intracellular calcium mobilization, and PKC–MAPK cascade activation that shapes gonadotrope transcriptional programs. This signaling axis integrates reproductive endocrine feedback and pulsatile stimulation dynamics, influencing cellular differentiation and hormone biosynthesis pathways. Dysregulated GNRHR expression or signaling is linked to disorders of reproductive axis function, making it a useful model for studying GPCR trafficking, desensitization, and endocrine signaling networks.

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

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

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