NK-1R CRISPR Activation Plasmid (h): sc-401004-ACT

TACR1 encodes the human neurokinin-1 receptor (NK-1R), a class A GPCR that binds the tachykinin neuropeptide substance P and couples primarily to Gq/11 signaling. NK-1R activation promotes phospholipase C–dependent inositol phosphate and calcium mobilization, with downstream engagement of PKC, MAPK/ERK, and transcriptional programs that modulate neuronal excitability, neurogenic inflammation, and smooth muscle contractility. TACR1 is expressed across central and peripheral nervous systems as well as immune and epithelial compartments, linking neuroimmune communication with barrier and vascular responses. Altered TACR1/NK-1R signaling has been associated with inflammatory and pain-related biology, airway and gastrointestinal dysregulation, and context-dependent changes in tumor microenvironment signaling, supporting mechanistic studies in disease-relevant cell models.

NK-1R CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TACR1 expression without altering the underlying DNA sequence.

NK-1R CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TACR1 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 TACR1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NK-1R expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TACR1 locus and enabling the study of NK-1R-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NK-1R pathway restoration in tumor cells with silenced or reduced TACR1 expression.

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