PTN CRISPR Activation Plasmid (m): sc-422498-ACT

Mouse Ptn encodes pleiotrophin (PTN), a secreted heparin-binding growth factor that signals through heparan sulfate proteoglycans and receptor protein tyrosine phosphatases to regulate cell proliferation, survival, migration, and neurite outgrowth. PTN activity intersects with MAPK/ERK, PI3K/AKT, and cytoskeletal remodeling pathways that shape tissue patterning and plasticity during development and repair. In the nervous system and stromal microenvironments, altered PTN expression is associated with changes in angiogenesis, inflammation, and extracellular matrix interactions that influence disease-relevant phenotypes. These properties make Ptn a useful target for mechanistic studies of neurodevelopment, injury responses, and microenvironmental signaling in mouse models.

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

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

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