PPX CRISPR Activation Plasmid (h): sc-403212-ACT

PPP4C encodes the catalytic subunit of protein phosphatase 4 (also known as PPX), a serine/threonine phosphatase that regulates phosphorylation-dependent signaling networks controlling cell cycle progression, centrosome organization, microtubule dynamics, and DNA damage responses. Through association with regulatory subunits, PPX modulates substrate selectivity in pathways linked to chromatin remodeling and checkpoint control, influencing genome stability and replication stress tolerance. Altered PP4/PPP4C activity has been associated with dysregulated proliferation and impaired repair signaling, making it relevant to studies of tumor biology and other disorders characterized by defective cell-cycle and stress-response pathways.

PPX CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PPP4C expression without altering the underlying DNA sequence.

PPX CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PPP4C 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 PPP4C transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PPX expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PPP4C locus and enabling the study of PPX-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PPX pathway restoration in tumor cells with silenced or reduced PPP4C expression.

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