Glutathione Peroxidase 8/GPX8 CRISPR Activation Plasmid (h): sc-404847-ACT

Human GPX8 (Glutathione Peroxidase 8) is an endoplasmic reticulum–associated glutathione peroxidase that contributes to redox homeostasis by reducing peroxides and limiting oxidative protein damage within the secretory pathway. By shaping ER oxidative folding conditions and peroxide flux, GPX8 influences proteostasis, unfolded protein response signaling, and broader cellular oxidative stress pathways. Altered GPX8 expression or activity has been linked in the literature to dysregulated redox balance and inflammatory signaling, with reported relevance across contexts such as tumor biology and metabolic stress. As a result, GPX8 is frequently studied for its role in ROS-dependent signaling, ER stress adaptation, and redox-controlled cell state transitions.

Glutathione Peroxidase 8/GPX8 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GPX8 expression without altering the underlying DNA sequence.

Glutathione Peroxidase 8/GPX8 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GPX8 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 GPX8 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Glutathione Peroxidase 8/GPX8 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GPX8 locus and enabling the study of Glutathione Peroxidase 8/GPX8-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Glutathione Peroxidase 8/GPX8 pathway restoration in tumor cells with silenced or reduced GPX8 expression.

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