PMP22 CRISPR Activation Plasmid (m): sc-422323-ACT

Mouse Pmp22 encodes peripheral myelin protein 22 (PMP22), a tetraspan membrane glycoprotein enriched in Schwann cells and compact myelin of the peripheral nervous system. PMP22 contributes to myelin sheath formation and maintenance by influencing membrane organization, Schwann cell differentiation, and proteostasis pathways that control folding and trafficking of myelin proteins. Altered PMP22 dosage perturbs myelination dynamics and axon–glia interactions, making Pmp22 a key locus for studying peripheral neuropathy mechanisms, stress responses in myelinating glia, and myelin-related signaling networks. In mouse models, modulation of Pmp22 expression is commonly used to interrogate gene dosage effects on myelin stability, nerve conduction properties, and downstream transcriptional programs.

PMP22 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Pmp22 expression without altering the underlying DNA sequence.

PMP22 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Pmp22 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 Pmp22 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PMP22 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Pmp22 locus and enabling the study of PMP22-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PMP22 pathway restoration in tumor cells with silenced or reduced Pmp22 expression.

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