PP5 CRISPR Activation Plasmid (h): sc-405979-ACT

PPP5C encodes serine/threonine-protein phosphatase 5 (PP5), a TPR-domain phosphatase that integrates signaling through its association with HSP90 chaperone complexes and multiple client proteins. PP5 regulates phosphorylation-dependent control of stress responses, cell cycle progression, and DNA damage signaling, including modulation of pathways linked to ATM/ATR signaling and MAPK networks. Through dephosphorylation of key regulators, PPP5C influences glucocorticoid receptor function, checkpoint control, and proteostasis, making it a useful node for studying signal transduction fidelity. Dysregulated PPP5C activity or expression has been associated with altered proliferation and stress tolerance phenotypes reported across cancer and neurobiology contexts, supporting its relevance in mechanistic disease models.

PP5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PPP5C expression without altering the underlying DNA sequence.

PP5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PPP5C 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 PPP5C transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PP5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PPP5C locus and enabling the study of PP5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PP5 pathway restoration in tumor cells with silenced or reduced PPP5C expression.

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