Trx CRISPR Activation Plasmid (h): sc-400746-ACT

Human TXN encodes thioredoxin (Trx), a ubiquitously expressed oxidoreductase that maintains cellular redox homeostasis by catalyzing dithiol–disulfide exchange reactions and supporting antioxidant defense systems. Trx interfaces with the thioredoxin reductase/NADPH axis and modulates redox-sensitive signaling nodes that influence transcriptional regulation, protein folding, and apoptosis, including regulation of ASK1-mediated stress signaling. Through control of intracellular reactive oxygen species and redox state, TXN contributes to processes such as proliferation, inflammatory signaling, and cellular responses to hypoxia and DNA damage. Dysregulated TXN/Trx activity has been associated with altered oxidative stress handling and signaling phenotypes observed across cancer biology, metabolic dysfunction, and neuroinflammatory contexts.

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

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

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