Glutathione reductase CRISPR Activation Plasmid (h): sc-417499-ACT

GSR encodes glutathione reductase, a flavoprotein oxidoreductase that maintains cellular redox homeostasis by regenerating reduced glutathione (GSH) from oxidized glutathione (GSSG) using NADPH. This reaction is central to antioxidant defense, supporting detoxification of reactive oxygen species and preserving thiol redox balance in the cytosol and mitochondria. By sustaining GSH-dependent pathways such as glutathione peroxidase activity and xenobiotic metabolism, GSR influences mitochondrial function, protein folding, and protection from oxidative damage. Dysregulated glutathione recycling and elevated oxidative stress are linked to mechanisms implicated in anemia, neurodegeneration, metabolic dysfunction, and cancer biology, making GSR a useful node for studying redox-driven phenotypes.

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

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

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