CYP46 CRISPR Activation Plasmid (m): sc-419926-ACT

Mouse Cyp46a1 encodes cholesterol 24-hydroxylase (CYP46A1/CYP46), a brain-enriched cytochrome P450 enzyme that catalyzes conversion of cholesterol to 24S-hydroxycholesterol to support cholesterol turnover and efflux across the blood–brain barrier. This reaction links neuronal sterol homeostasis to broader lipid metabolic networks, including oxysterol signaling and regulation of cholesterol-dependent membrane composition that influences synaptic function. Altered CYP46A1 activity is associated with disruptions in brain cholesterol balance and has been studied in the context of neurodegenerative and cognitive phenotypes, where sterol dysregulation intersects with neuroinflammation and proteostasis pathways.

CYP46 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Cyp46a1 expression without altering the underlying DNA sequence.

CYP46 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Cyp46a1 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 Cyp46a1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CYP46 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Cyp46a1 locus and enabling the study of CYP46-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CYP46 pathway restoration in tumor cells with silenced or reduced Cyp46a1 expression.

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