PME-1 CRISPR Activation Plasmid (h): sc-402446-ACT

Human PPME1 encodes protein phosphatase methylesterase-1 (PME-1), an enzyme that hydrolyzes the C-terminal methyl ester of PP2A catalytic subunits and thereby modulates PP2A holoenzyme assembly, localization, and substrate selection. By regulating PP2A methylation status, PME-1 influences phosphorylation-dependent control of cell cycle progression, DNA damage signaling, and stress-responsive pathways including MAPK and PI3K/AKT network outputs. Altered PP2A regulation is broadly relevant to mechanisms of aberrant kinase signaling and proteostasis that are frequently studied in cancer biology and neurodegeneration. PPME1 is therefore a useful node for dissecting how phosphatase tuning reshapes signaling dynamics and cellular phenotypes under physiological and disease-associated conditions.

PME-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PPME1 expression without altering the underlying DNA sequence.

PME-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PPME1 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 PPME1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PME-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PPME1 locus and enabling the study of PME-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PME-1 pathway restoration in tumor cells with silenced or reduced PPME1 expression.

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