PP2A-B56-δ CRISPR Activation Plasmid (h): sc-403020-ACT

PPP2R5D encodes the B56δ regulatory subunit of protein phosphatase 2A (PP2A), a major serine/threonine phosphatase that shapes phospho-signaling by directing substrate selectivity and subcellular localization of the PP2A holoenzyme. PP2A-B56δ contributes to control of cell-cycle progression, DNA damage signaling, and mitotic checkpoint fidelity by modulating phosphorylation states of key pathway nodes, thereby influencing proliferation and stress responses. Through PP2A-dependent tuning of kinase-driven networks such as MAPK/ERK and PI3K/AKT, PPP2R5D helps maintain signaling homeostasis and transcriptional programs linked to differentiation and survival. Dysregulation or mutation of PP2A regulatory subunits, including PPP2R5D, is associated with altered phosphorylation landscapes relevant to cancer biology and neurodevelopmental phenotypes, supporting its use as a mechanistic target in pathway-focused studies.

PP2A-B56-δ CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PPP2R5D expression without altering the underlying DNA sequence.

PP2A-B56-δ CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PPP2R5D 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 PPP2R5D transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PP2A-B56-δ expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PPP2R5D locus and enabling the study of PP2A-B56-δ-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PP2A-B56-δ pathway restoration in tumor cells with silenced or reduced PPP2R5D expression.

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