Peptide YY CRISPR Activation Plasmid (h): sc-403446-ACT

Human PYY encodes peptide YY, a gut-derived endocrine peptide released primarily from enteroendocrine L cells in response to nutrient intake. Peptide YY engages neuroendocrine signaling through Y-family G protein–coupled receptors, modulating cAMP-linked pathways that influence appetite regulation, gastrointestinal motility, and pancreatic secretion. In peripheral tissues and the central nervous system, PYY contributes to coordinated metabolic homeostasis and communication along the gut–brain axis. Altered PYY expression or secretion has been associated with dysregulated energy balance and metabolic phenotypes, supporting its use as a molecular readout in studies of obesity, type 2 diabetes–related physiology, and gastrointestinal function.

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

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

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