AKR7A2 CRISPR Activation Plasmid (h): sc-403377-ACT

AKR7A2 encodes a cytosolic aldo-keto reductase that catalyzes NADPH-dependent reduction of reactive aldehydes and ketones, supporting cellular defense against electrophilic and oxidative stress. By detoxifying lipid peroxidation-derived aldehydes and xenobiotic metabolites, AKR7A2 contributes to redox homeostasis and maintenance of cellular proteome integrity. Its activity intersects with aldehyde detoxification networks and broader stress-response pathways that shape susceptibility to toxicant exposure and inflammatory signaling. Altered regulation of AKR7A2 has been investigated in contexts where carbonyl stress and xenobiotic metabolism influence disease-relevant phenotypes, including hepatic dysfunction and tumor-associated oxidative stress states.

AKR7A2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous AKR7A2 expression without altering the underlying DNA sequence.

AKR7A2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the AKR7A2 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 AKR7A2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous AKR7A2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native AKR7A2 locus and enabling the study of AKR7A2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of AKR7A2 pathway restoration in tumor cells with silenced or reduced AKR7A2 expression.

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