NTR1 CRISPR Activation Plasmid (h): sc-402552-ACT

Neurotensin receptor 1 (NTSR1/NTR1) is a class A GPCR that couples primarily to Gq/11 and can also engage β-arrestin–dependent signaling, linking neurotensin binding to phospholipase C activation, intracellular Ca2+ mobilization, and MAPK/ERK pathway output. Through these cascades, NTR1 influences neurotransmission, endocrine secretion, smooth muscle contractility, and regulation of cellular proliferation and survival programs. NTSR1 expression and signaling have been investigated in contexts of altered neuronal excitability, inflammatory signaling, and oncogenic pathway crosstalk, where receptor-driven MAPK and PI3K/AKT activity can modulate cell-state transitions. These features make NTSR1 a useful node for studying GPCR-driven transcriptional programs, receptor desensitization/internalization, and pathway rewiring in disease-relevant models.

NTR1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NTSR1 expression without altering the underlying DNA sequence.

NTR1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NTSR1 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 NTSR1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NTR1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NTSR1 locus and enabling the study of NTR1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NTR1 pathway restoration in tumor cells with silenced or reduced NTSR1 expression.

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