AKR1B10 CRISPR Activation Plasmid (h): sc-403649-ACT

AKR1B10 encodes a human NADPH-dependent aldo-keto reductase that reduces reactive carbonyls and lipid peroxidation–derived aldehydes, contributing to cellular redox homeostasis and detoxification. By modulating retinoid and lipid metabolism and buffering oxidative stress, AKR1B10 influences proliferation, differentiation, and inflammatory signaling programs. Dysregulated AKR1B10 expression has been reported across multiple tumor contexts and in metabolic and oxidative stress–associated states, where altered carbonyl handling can reshape cellular fitness. As a result, AKR1B10 is frequently studied as a node linking xenobiotic response, lipid remodeling, and stress-adaptation pathways.

AKR1B10 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous AKR1B10 expression without altering the underlying DNA sequence.

AKR1B10 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the AKR1B10 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 AKR1B10 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous AKR1B10 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native AKR1B10 locus and enabling the study of AKR1B10-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of AKR1B10 pathway restoration in tumor cells with silenced or reduced AKR1B10 expression.

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