PLP CRISPR Activation Plasmid (h): sc-401663-ACT

PLP1 encodes proteolipid protein 1 (PLP), a major structural component of compact myelin in the central nervous system that supports oligodendrocyte membrane architecture and efficient axonal conduction. PLP participates in myelin sheath assembly and maintenance through lipid-rich membrane organization and myelin biogenesis pathways that govern white matter integrity. Altered PLP1 dosage or sequence disrupts myelination and oligodendrocyte function and is associated with inherited hypomyelinating leukodystrophies and related neurodevelopmental and neurodegenerative phenotypes. As a result, PLP1 is frequently studied in models of oligodendrocyte differentiation, axon–glia interactions, and mechanisms of myelin stability.

PLP CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PLP1 expression without altering the underlying DNA sequence.

PLP CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PLP1 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 PLP1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PLP expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PLP1 locus and enabling the study of PLP-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PLP pathway restoration in tumor cells with silenced or reduced PLP1 expression.

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