Glutathione Peroxidase 4/GPX4 CRISPR Activation Plasmid (h): sc-401558-ACT

GPX4 (glutathione peroxidase 4) encodes a selenium-dependent phospholipid hydroperoxidase that reduces membrane lipid hydroperoxides using glutathione, maintaining redox homeostasis and preserving membrane integrity. As a central suppressor of ferroptosis, GPX4 integrates with glutathione metabolism, lipid peroxidation control, and antioxidant defense pathways, influencing mitochondrial and ER-associated oxidative stress responses. Dysregulated GPX4 activity has been linked to altered sensitivity to oxidative injury and ferroptotic cell death in contexts including cancer biology, neurodegeneration, and inflammatory tissue damage. Its expression and enzymatic capacity are therefore widely studied in models of lipid ROS accumulation, iron-dependent cell death, and stress-adaptation signaling.

Glutathione Peroxidase 4/GPX4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GPX4 expression without altering the underlying DNA sequence.

Glutathione Peroxidase 4/GPX4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GPX4 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 GPX4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Glutathione Peroxidase 4/GPX4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GPX4 locus and enabling the study of Glutathione Peroxidase 4/GPX4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Glutathione Peroxidase 4/GPX4 pathway restoration in tumor cells with silenced or reduced GPX4 expression.

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