CYP7A1 CRISPR Activation Plasmid (h): sc-401001-ACT

Human CYP7A1 encodes cholesterol 7α-hydroxylase, a liver-enriched cytochrome P450 enzyme that catalyzes the first and rate-limiting step of the classic bile acid synthesis pathway from cholesterol. By controlling conversion of cholesterol to primary bile acids, CYP7A1 influences cholesterol homeostasis, enterohepatic signaling, and feedback regulation via nuclear receptors such as FXR and downstream metabolic gene programs. Altered CYP7A1 activity has been linked to dysregulated bile acid pools, hypercholesterolemia susceptibility, and metabolic phenotypes affecting lipid absorption and hepatic lipid handling. As a central node in sterol and bile acid metabolism, CYP7A1 is widely studied in hepatocyte biology, metabolic regulation, and pathway cross-talk with inflammation and xenobiotic responses.

CYP7A1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CYP7A1 expression without altering the underlying DNA sequence.

CYP7A1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CYP7A1 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 CYP7A1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CYP7A1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CYP7A1 locus and enabling the study of CYP7A1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CYP7A1 pathway restoration in tumor cells with silenced or reduced CYP7A1 expression.

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