Grx1 CRISPR Activation Plasmid (h): sc-418056-ACT

Human GLRX encodes glutaredoxin-1 (Grx1), a cytosolic thiol-disulfide oxidoreductase that catalyzes deglutathionylation and helps maintain cellular redox homeostasis. By regulating protein S-glutathionylation, Grx1 influences signaling and stress-response pathways linked to glutathione metabolism, antioxidant defenses, and redox-sensitive control of apoptosis and inflammation. Grx1 activity impacts mitochondrial function, proteostasis, and responses to reactive oxygen and nitrogen species, integrating with broader oxidative stress programs. Dysregulated GLRX/Grx1 function has been associated with redox imbalance observed across multiple disease-relevant contexts, including inflammatory and metabolic stress phenotypes and altered cellular resilience to oxidative injury.

Grx1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GLRX expression without altering the underlying DNA sequence.

Grx1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GLRX 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 GLRX transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Grx1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GLRX locus and enabling the study of Grx1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Grx1 pathway restoration in tumor cells with silenced or reduced GLRX expression.

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