MC3-R CRISPR Activation Plasmid (h): sc-403341-ACT

MC3R encodes melanocortin-3 receptor (MC3-R), a G protein–coupled receptor that responds to melanocortin peptides to regulate energy homeostasis, nutrient partitioning, and neuroendocrine signaling. Upon ligand engagement, MC3-R predominantly couples to Gs to elevate cAMP and activate PKA/CREB-dependent transcriptional programs, with downstream effects on hypothalamic circuitry and peripheral metabolism. MC3R activity is linked to pathways integrating appetite, circadian and feeding-related behaviors, and inflammatory cues within metabolic tissues. Genetic and functional perturbations of MC3R have been associated with body weight regulation phenotypes and metabolic dysfunction, supporting its relevance in studies of obesity biology and related cardiometabolic traits.

MC3-R CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MC3R expression without altering the underlying DNA sequence.

MC3-R CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MC3R 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 MC3R transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MC3-R expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MC3R locus and enabling the study of MC3-R-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MC3-R pathway restoration in tumor cells with silenced or reduced MC3R expression.

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