Glutathione Peroxidase 3/GPX3 CRISPR Activation Plasmid (h): sc-402413-ACT

Human GPX3 encodes glutathione peroxidase 3, a secreted selenoprotein that reduces hydrogen peroxide and lipid hydroperoxides using glutathione, thereby limiting extracellular oxidative damage. GPX3 contributes to redox homeostasis, modulation of inflammatory signaling, and protection of membrane and matrix components from reactive oxygen species, intersecting with oxidative stress response networks and glutathione metabolism. Altered GPX3 expression has been reported in contexts associated with dysregulated redox balance, including cancer biology, metabolic stress, and vascular or tissue remodeling processes, where extracellular antioxidant capacity can influence signaling and cell–microenvironment interactions. These features make GPX3 a useful target for studying redox-regulated pathways, oxidative stress phenotypes, and secreted antioxidant function in human cell models.

Glutathione Peroxidase 3/GPX3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GPX3 expression without altering the underlying DNA sequence.

Glutathione Peroxidase 3/GPX3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GPX3 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 GPX3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Glutathione Peroxidase 3/GPX3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GPX3 locus and enabling the study of Glutathione Peroxidase 3/GPX3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Glutathione Peroxidase 3/GPX3 pathway restoration in tumor cells with silenced or reduced GPX3 expression.

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