GSTM1 CRISPR Activation Plasmid (h): sc-418201-ACT

Glutathione S-transferase mu 1 (GSTM1) is a cytosolic phase II detoxification enzyme that catalyzes glutathione conjugation to electrophilic xenobiotics and endogenous lipid peroxidation products, supporting redox homeostasis and cellular defense against oxidative stress. GSTM1 activity intersects with glutathione metabolism, reactive oxygen species buffering, and chemical carcinogenesis pathways by facilitating the clearance of reactive intermediates. Genetic variation and deletion polymorphisms in GSTM1 are widely studied for their impact on xenobiotic handling and susceptibility to toxicant-induced cellular injury. Altered GSTM1 expression has been associated with differences in inflammatory signaling and stress-response phenotypes relevant to cancer biology, pulmonary disease, and drug metabolism research.

GSTM1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GSTM1 expression without altering the underlying DNA sequence.

GSTM1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GSTM1 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 GSTM1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GSTM1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GSTM1 locus and enabling the study of GSTM1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GSTM1 pathway restoration in tumor cells with silenced or reduced GSTM1 expression.

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