ghrelin CRISPR Activation Plasmid (m): sc-425542-ACT

Mouse Ghrl encodes ghrelin, a secreted peptide hormone primarily produced by the stomach that signals through the growth hormone secretagogue receptor to coordinate energy balance and neuroendocrine function. Ghrelin regulates appetite, adiposity, glucose homeostasis, gastrointestinal motility, and growth hormone release, integrating peripheral metabolic status with hypothalamic and pituitary signaling networks. Downstream processes intersect with AMPK-dependent nutrient sensing and circuits controlling reward, stress responses, and inflammation. Altered ghrelin signaling has been associated with obesity and metabolic syndrome phenotypes, as well as cachexia, insulin resistance, and neurobehavioral changes in experimental models.

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

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

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