NPY CRISPR Activation Plasmid (h): sc-401098-ACT

Human NPY encodes neuropeptide Y, a secreted peptide neurotransmitter that signals primarily through Y-family G protein–coupled receptors to regulate neuronal excitability, synaptic transmission, and neuroendocrine outputs. NPY is a key modulator of energy homeostasis, feeding behavior, circadian and stress responses, and autonomic control, with downstream effects on cAMP and calcium-dependent signaling in neural and peripheral tissues. In addition to central nervous system functions, NPY influences vascular tone and immune cell activity, linking neuroimmune and cardiometabolic pathways. Dysregulated NPY signaling has been associated with neuropsychiatric phenotypes, obesity-related traits, and cardiovascular risk biology, making it a widely used marker and functional node for mechanistic studies.

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

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

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