Glutathione Peroxidase 2/GPX2 CRISPR Activation Plasmid (h): sc-406827-ACT

Human GPX2 (glutathione peroxidase 2) is a selenium-dependent antioxidant enzyme that reduces hydrogen peroxide and lipid hydroperoxides using glutathione, helping maintain redox homeostasis at epithelial barriers. By limiting oxidative damage, GPX2 influences ROS-sensitive signaling, including NRF2-driven stress responses, inflammatory pathways, and cellular decisions around proliferation and apoptosis. GPX2 activity is closely linked to glutathione metabolism and broader detoxification processes that shape mitochondrial function and membrane lipid integrity. Dysregulated GPX2 expression has been associated with oxidative stress phenotypes and inflammation-linked pathologies, and it is frequently studied in contexts such as gastrointestinal biology, epithelial differentiation, and tumor-associated redox adaptation.

Glutathione Peroxidase 2/GPX2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GPX2 expression without altering the underlying DNA sequence.

Glutathione Peroxidase 2/GPX2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GPX2 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 GPX2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Glutathione Peroxidase 2/GPX2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GPX2 locus and enabling the study of Glutathione Peroxidase 2/GPX2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Glutathione Peroxidase 2/GPX2 pathway restoration in tumor cells with silenced or reduced GPX2 expression.

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