PrP CRISPR Activation Plasmid (h): sc-401061-ACT

PRNP encodes the cellular prion protein (PrP), a glycosylphosphatidylinositol-anchored membrane glycoprotein enriched at the neuronal surface and within lipid rafts. PrP participates in proteostasis and membrane-associated signaling processes, including endocytic trafficking, redox homeostasis, and modulation of synaptic function through interactions with extracellular matrix components and cell-adhesion partners. Misfolding of PrP into protease-resistant conformers underlies transmissible spongiform encephalopathies and is linked to neurodegeneration characterized by protein aggregation, altered cellular stress responses, and progressive neuronal dysfunction. PRNP regulation is therefore relevant for studying pathways governing protein folding quality control, neuroinflammatory signaling, and neuronal survival mechanisms.

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

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

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