AKR1C14 CRISPR Activation Plasmid (m): sc-430591-ACT

Akr1c14 encodes AKR1C14, a mouse aldo-keto reductase that catalyzes NADPH-dependent reduction of carbonyl-containing substrates, including endogenous steroids and lipid aldehydes. By modulating local steroid hormone availability and detoxifying reactive carbonyl species, AKR1C14 contributes to redox homeostasis and metabolic signaling in tissues such as liver and reproductive organs. This enzymatic activity intersects with pathways linked to xenobiotic metabolism, oxidative stress responses, and regulation of nuclear receptor signaling. Altered aldo-keto reductase activity is frequently studied in the context of metabolic dysfunction, inflammation, and hormone-dependent physiology, making Akr1c14 a useful node for mechanistic investigations in mouse models.

AKR1C14 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Akr1c14 expression without altering the underlying DNA sequence.

AKR1C14 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Akr1c14 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 Akr1c14 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous AKR1C14 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Akr1c14 locus and enabling the study of AKR1C14-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of AKR1C14 pathway restoration in tumor cells with silenced or reduced Akr1c14 expression.

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