GADD 34 CRISPR Activation Plasmid (h): sc-400595-ACT

PPP1R15A encodes GADD34, a regulatory subunit of protein phosphatase 1 that promotes dephosphorylation of eIF2α to terminate the integrated stress response and restore global protein synthesis following cellular stress. Induced downstream of ATF4/CHOP signaling, GADD34 functions as a key feedback node within ER stress and unfolded protein response pathways, shaping proteostasis, apoptosis sensitivity, and recovery from translational arrest. Through its influence on stress-adaptive signaling, PPP1R15A is studied in contexts including inflammation, metabolic stress, neurodegeneration, and tumor cell survival, where dysregulated stress responses can contribute to disease phenotypes. Its activity also intersects with oxidative stress and DNA damage–associated programs, making it a useful target for mechanistic interrogation of stress signaling networks.

GADD 34 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PPP1R15A expression without altering the underlying DNA sequence.

GADD 34 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PPP1R15A 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 PPP1R15A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GADD 34 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PPP1R15A locus and enabling the study of GADD 34-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GADD 34 pathway restoration in tumor cells with silenced or reduced PPP1R15A expression.

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