MCH-1R CRISPR Activation Plasmid (h): sc-402512-ACT

MCHR1 encodes melanin-concentrating hormone receptor 1 (MCH-1R), a class A GPCR that binds MCH to modulate neuronal signaling involved in energy homeostasis, feeding behavior, and neuroendocrine regulation. Upon activation, MCH-1R primarily couples to G proteins to influence second-messenger signaling, including cAMP and calcium-dependent pathways, and can engage MAPK/ERK signaling to shape downstream transcriptional responses. MCHR1 expression in hypothalamic and limbic circuits links this receptor to regulation of appetite, arousal, and stress-responsive behaviors. Dysregulated MCH-1R signaling has been studied in the context of obesity and metabolic imbalance, as well as neuropsychiatric phenotypes where altered neuromodulatory tone may contribute to disease-associated circuitry changes.

MCH-1R CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MCHR1 expression without altering the underlying DNA sequence.

MCH-1R CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MCHR1 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 MCHR1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MCH-1R expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MCHR1 locus and enabling the study of MCH-1R-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MCH-1R pathway restoration in tumor cells with silenced or reduced MCHR1 expression.

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