PPP1R3 CRISPR Activation Plasmid (h): sc-405135-ACT

PPP1R3A encodes PPP1R3, a glycogen-targeting regulatory subunit of protein phosphatase 1 that helps localize catalytic PP1 to glycogen particles and coordinate dephosphorylation of key metabolic enzymes. By modulating glycogen synthase and phosphorylase activity, PPP1R3A contributes to glucose storage dynamics and integrates insulin-responsive signaling with carbohydrate metabolism in skeletal muscle and other insulin-sensitive tissues. Altered PPP1R3A expression or regulation has been linked to dysregulated glycogen handling and metabolic phenotypes, making it relevant for studies of insulin signaling, energy homeostasis, and tissue-specific metabolic adaptation. As a scaffold-like PP1 regulator, PPP1R3A also serves as a tractable node to dissect phosphatase targeting mechanisms within broader phosphorylation networks.

PPP1R3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PPP1R3A expression without altering the underlying DNA sequence.

PPP1R3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PPP1R3A 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 PPP1R3A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PPP1R3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PPP1R3A locus and enabling the study of PPP1R3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PPP1R3 pathway restoration in tumor cells with silenced or reduced PPP1R3A expression.

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