GHR CRISPR Activation Plasmid (m): sc-420549-ACT

Mouse Ghr encodes the growth hormone receptor (GHR), a transmembrane cytokine receptor that binds growth hormone to regulate somatic growth, metabolism, and tissue homeostasis. Ligand engagement promotes receptor dimerization and JAK2 activation, driving downstream STAT5 signaling as well as cross-talk with PI3K–AKT and MAPK/ERK pathways to control transcriptional programs for proliferation, differentiation, and nutrient utilization. GHR activity shapes hepatic IGF-1 production and influences adipose, muscle, and bone physiology, making it central to endocrine regulation of growth and energy balance. Dysregulated GHR signaling has been associated with altered growth phenotypes and metabolic dysfunction, and it is frequently studied in contexts of developmental biology, aging, and endocrine pathway remodeling.

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

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

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