PP2A-B56-γ CRISPR Activation Plasmid (h): sc-402726-ACT

PPP2R5C encodes the B56γ regulatory subunit of protein phosphatase 2A (PP2A), a major serine/threonine phosphatase that directs catalytic activity toward specific substrates and subcellular locations. PP2A-B56γ contributes to control of cell-cycle progression, DNA damage signaling, mitotic checkpoint fidelity, and signal transduction through pathways such as PI3K–AKT, MAPK, and Wnt/β-catenin by coordinating dephosphorylation of key regulators. Altered PPP2R5C expression or PP2A holoenzyme composition can shift phosphorylation networks that influence proliferation, genome stability, and stress responses. These properties make PP2A-B56γ a useful node for studying phosphatase-guided pathway insulation and mechanisms linking dysregulated dephosphorylation to disease-associated phenotypes.

PP2A-B56-γ CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PPP2R5C expression without altering the underlying DNA sequence.

PP2A-B56-γ CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PPP2R5C 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 PPP2R5C transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PP2A-B56-γ expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PPP2R5C locus and enabling the study of PP2A-B56-γ-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PP2A-B56-γ pathway restoration in tumor cells with silenced or reduced PPP2R5C expression.

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