NPY2-R CRISPR Activation Plasmid (h): sc-417895-ACT

NPY2R encodes the human neuropeptide Y receptor Y2 (NPY2-R), a Gi/o-coupled GPCR that binds neuropeptide Y and peptide YY to modulate cAMP signaling, ion channel activity, and downstream MAPK/ERK pathway dynamics. NPY2-R is broadly implicated in neuroendocrine regulation, shaping synaptic transmission, stress responsivity, and appetite-related circuits, and it also contributes to vascular and gastrointestinal physiology. Through inhibitory neurotransmission and presynaptic control of peptide release, NPY2R influences cellular excitability and metabolic homeostasis. Dysregulated NPY2R signaling has been associated with phenotypes relevant to energy balance, mood and stress pathways, and inflammatory or autonomic processes, making it a useful target for mechanistic studies in neuronal and peripheral cell models.

NPY2-R CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NPY2R expression without altering the underlying DNA sequence.

NPY2-R CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NPY2R 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 NPY2R transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NPY2-R expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NPY2R locus and enabling the study of NPY2-R-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NPY2-R pathway restoration in tumor cells with silenced or reduced NPY2R expression.

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