PPP1R12C CRISPR Activation Plasmid (h): sc-412332-ACT

PPP1R12C encodes MYPT3, a regulatory subunit of myosin phosphatase that targets protein phosphatase 1 (PP1) to myosin II and related substrates to modulate actomyosin contractility. By shaping myosin light chain phosphorylation dynamics, PPP1R12C contributes to cytoskeletal organization, cell adhesion, motility, and mechanotransduction pathways that influence tissue architecture and barrier function. Altered regulation of PP1-dependent dephosphorylation has been linked to dysregulated cell migration and contractile signaling, processes commonly implicated in cardiovascular and cancer biology. As a node connecting phosphatase targeting with RhoA/ROCK-driven contractility, PPP1R12C is relevant for studies of phosphorylation signaling networks and cytoskeleton-dependent phenotypes.

PPP1R12C CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PPP1R12C expression without altering the underlying DNA sequence.

PPP1R12C CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PPP1R12C 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 PPP1R12C transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PPP1R12C expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PPP1R12C locus and enabling the study of PPP1R12C-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PPP1R12C pathway restoration in tumor cells with silenced or reduced PPP1R12C expression.

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