GSTT1 CRISPR Activation Plasmid (h): sc-408735-ACT

Human GSTT1 encodes glutathione S-transferase theta 1, a phase II detoxification enzyme that catalyzes the conjugation of reduced glutathione to electrophilic compounds, supporting cellular redox homeostasis and defense against xenobiotics and oxidative stress. GSTT1 activity contributes to glutathione-dependent metabolism of environmental chemicals and endogenous lipid peroxidation products, intersecting with pathways that modulate reactive oxygen species handling and stress-response signaling. Genetic variation or deletion of GSTT1 has been associated with altered susceptibility to toxicant-induced damage and interindividual differences in chemical metabolism, making it a frequent variable in studies of carcinogenesis risk modifiers, inflammatory processes, and pharmacogenomic responses. As a cytosolic enzyme with broad substrate scope, GSTT1 is also used as a model to interrogate detoxification capacity and compensatory regulation among GST family members.

GSTT1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GSTT1 expression without altering the underlying DNA sequence.

GSTT1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GSTT1 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 GSTT1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GSTT1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GSTT1 locus and enabling the study of GSTT1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GSTT1 pathway restoration in tumor cells with silenced or reduced GSTT1 expression.

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