NMUR1 CRISPR Activation Plasmid (h): sc-405595-ACT

Neuromedin U receptor 1 (NMUR1) is a class A G protein–coupled receptor that binds the neuropeptide neuromedin U and couples primarily to Gq/11 signaling to elevate intracellular Ca²⁺ and activate downstream MAPK and PKC-dependent transcriptional programs. NMUR1 is expressed in immune and peripheral tissues where it modulates inflammatory signaling, cytokine production, smooth muscle contractility, and neuroendocrine communication. Through these pathways, NMUR1 has been studied in mechanisms linking neuroimmune regulation to airway and gastrointestinal physiology, as well as systemic metabolic homeostasis. Dysregulated NMUR1 signaling has been associated in research settings with inflammatory disorders and altered metabolic phenotypes, making it a useful target for pathway dissection in relevant cellular models.

NMUR1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NMUR1 expression without altering the underlying DNA sequence.

NMUR1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NMUR1 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 NMUR1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NMUR1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NMUR1 locus and enabling the study of NMUR1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NMUR1 pathway restoration in tumor cells with silenced or reduced NMUR1 expression.

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