TrxR2 CRISPR Activation Plasmid (h): sc-402671-ACT

Human TXNRD2 encodes mitochondrial thioredoxin reductase 2 (TrxR2), a selenoenzyme that maintains thioredoxin-2 in its reduced state to support mitochondrial redox homeostasis. TrxR2 integrates NADPH-dependent antioxidant defense with control of reactive oxygen species, mitochondrial protein thiol status, and redox-sensitive signaling that influences apoptosis and metabolic adaptation. Through its role in thioredoxin and peroxiredoxin systems, TXNRD2 contributes to oxidative phosphorylation integrity and cellular responses to oxidative stress. Dysregulation of mitochondrial redox control involving TXNRD2 has been associated with contexts of altered metabolism, neurodegeneration, and cancer biology, motivating mechanistic studies of redox-dependent phenotypes.

TrxR2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TXNRD2 expression without altering the underlying DNA sequence.

TrxR2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TXNRD2 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 TXNRD2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TrxR2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TXNRD2 locus and enabling the study of TrxR2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TrxR2 pathway restoration in tumor cells with silenced or reduced TXNRD2 expression.

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