Glutathione Peroxidase 5/GPX5 CRISPR Activation Plasmid (h): sc-405279-ACT

Human GPX5 encodes glutathione peroxidase 5, a selenium-dependent antioxidant enzyme that reduces hydrogen peroxide and lipid hydroperoxides using glutathione, thereby limiting oxidative damage to proteins and membranes. By modulating cellular redox homeostasis, GPX5 can influence ROS-sensitive signaling networks and oxidative stress responses that intersect with inflammation, mitochondrial function, and apoptotic pathways. Altered peroxide detoxification capacity is broadly relevant to disease mechanisms in which oxidative injury contributes to tissue dysfunction, including cancer biology, neurodegeneration, and cardiometabolic stress. GPX5 is also studied in reproductive and secretory tract physiology where redox regulation supports cellular viability and protein quality control.

Glutathione Peroxidase 5/GPX5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GPX5 expression without altering the underlying DNA sequence.

Glutathione Peroxidase 5/GPX5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GPX5 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 GPX5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Glutathione Peroxidase 5/GPX5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GPX5 locus and enabling the study of Glutathione Peroxidase 5/GPX5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Glutathione Peroxidase 5/GPX5 pathway restoration in tumor cells with silenced or reduced GPX5 expression.

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