Carbonyl reductase 1 CRISPR Activation Plasmid (h): sc-402755-ACT

Human CBR1 encodes carbonyl reductase 1, a cytosolic NADPH-dependent oxidoreductase that converts a wide range of reactive aldehydes and ketones to less reactive alcohols. By processing lipid peroxidation-derived carbonyls and other electrophiles, CBR1 contributes to cellular redox homeostasis, oxidative stress responses, and metabolism of endogenous and xenobiotic carbonyl compounds. Altered CBR1 activity has been linked to variation in carbonyl detoxification capacity and redox imbalance observed across cancer biology, inflammation, and metabolic dysfunction, making it relevant for pathway-level studies of oxidative damage and adaptive stress signaling. Its enzymatic position connects to broader aldehyde detoxification and NADPH-utilizing networks that influence proteostasis and cellular survival under stress.

Carbonyl reductase 1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CBR1 expression without altering the underlying DNA sequence.

Carbonyl reductase 1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CBR1 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 CBR1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Carbonyl reductase 1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CBR1 locus and enabling the study of Carbonyl reductase 1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Carbonyl reductase 1 pathway restoration in tumor cells with silenced or reduced CBR1 expression.

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