PPP1R15B CRISPR Activation Plasmid (h): sc-408988-ACT

PPP1R15B (also known as CReP) encodes a regulatory subunit that targets protein phosphatase 1 (PP1) to dephosphorylate eIF2α, acting as a key counterbalance to stress-induced translational repression. By promoting eIF2α dephosphorylation, PPP1R15B helps restore global protein synthesis after perturbations and modulates integrated stress response signaling that intersects with ER stress, proteostasis, and cell survival programs. This PP1-directed control influences downstream transcriptional and metabolic adaptation pathways governed by ATF4/CHOP-dependent networks. Dysregulation of eIF2α phosphorylation dynamics and ISR tuning has been implicated in diverse contexts including neurodegeneration, metabolic stress, and cancer cell adaptation, making PPP1R15B a useful node for mechanistic studies of stress resilience.

PPP1R15B CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PPP1R15B expression without altering the underlying DNA sequence.

PPP1R15B CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PPP1R15B 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 PPP1R15B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PPP1R15B expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PPP1R15B locus and enabling the study of PPP1R15B-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PPP1R15B pathway restoration in tumor cells with silenced or reduced PPP1R15B expression.

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