Peripherin CRISPR Activation Plasmid (h): sc-401335-ACT

PRPH encodes peripherin, a type III intermediate filament protein that is enriched in peripheral neurons and contributes to cytoskeletal architecture and axonal integrity. Peripherin participates in intermediate filament assembly and remodeling that supports neurite extension, axonal transport, and responses to cellular stress during neuronal differentiation and regeneration. Its expression is commonly used as a marker of peripheral nervous system development and injury-associated reprogramming. Dysregulated PRPH expression and filament organization have been linked to neurodegenerative processes and motor neuron vulnerability, supporting its relevance in mechanistic studies of neuronal dysfunction.

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

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

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