GSTO1 CRISPR Activation Plasmid (h): sc-404107-ACT

Human GSTO1 encodes glutathione S-transferase omega-1, an atypical GST family enzyme that catalyzes thioltransferase and deglutathionylation reactions important for maintaining cellular redox balance. By regulating S-glutathionylation status of protein thiols, GSTO1 contributes to antioxidant defense, proteostasis, and stress-responsive signaling that interfaces with glutathione metabolism and detoxification pathways. Altered GSTO1 activity and genetic variation have been associated with susceptibility and progression in disorders linked to oxidative stress and inflammation, including neurodegenerative and cardiometabolic phenotypes, as well as aspects of tumor biology. These properties make GSTO1 a useful node for interrogating redox-regulated signaling, mitochondrial and ER stress responses, and the impact of glutathione-dependent chemistry on cell fate decisions.

GSTO1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GSTO1 expression without altering the underlying DNA sequence.

GSTO1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GSTO1 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 GSTO1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GSTO1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GSTO1 locus and enabling the study of GSTO1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GSTO1 pathway restoration in tumor cells with silenced or reduced GSTO1 expression.

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