GnRH I CRISPR Activation Plasmid (m): sc-420625-ACT

Mouse Gnrh1 encodes gonadotropin-releasing hormone I (GnRH I), a hypothalamic neuropeptide that orchestrates reproductive endocrine function by stimulating pituitary gonadotrophs to secrete luteinizing hormone and follicle-stimulating hormone. Pulsatile GnRH I signaling engages GnRH receptor–dependent Gq/11 pathways, driving PLCβ activation, intracellular Ca2+ mobilization, PKC/MAPK cascades, and transcriptional programs that regulate gonadotropin synthesis and release. Through integration with kisspeptin, sex steroid feedback, and circadian/metabolic inputs, Gnrh1 contributes to puberty timing, fertility, and cyclic reproductive physiology. Dysregulation of GnRH I production or signaling is relevant to experimental models of hypogonadotropic hypogonadism, pubertal disorders, and neuroendocrine perturbations affecting reproductive axis homeostasis.

GnRH I CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Gnrh1 expression without altering the underlying DNA sequence.

GnRH I CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Gnrh1 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 Gnrh1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GnRH I expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Gnrh1 locus and enabling the study of GnRH I-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GnRH I pathway restoration in tumor cells with silenced or reduced Gnrh1 expression.

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