CYP8B1 CRISPR Activation Plasmid (h): sc-403383-ACT

Human CYP8B1 encodes sterol 12α-hydroxylase, a microsomal cytochrome P450 enzyme that defines bile acid composition by catalyzing a key hydroxylation step required for cholic acid synthesis. Through its role in the classic bile acid biosynthesis pathway, CYP8B1 influences the cholic acid/chenodeoxycholic acid ratio, thereby modulating hepatic cholesterol turnover, intestinal lipid absorption, and bile acid–mediated signaling. CYP8B1 activity is functionally linked to metabolic homeostasis via crosstalk with nuclear receptor pathways such as FXR and related transcriptional programs governing bile acid transport and feedback regulation. Altered CYP8B1 expression or bile acid profiles has been associated with dyslipidemia, insulin resistance, nonalcoholic fatty liver disease, and cholestatic liver phenotypes, making it a useful node for mechanistic studies of hepatobiliary metabolism.

CYP8B1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CYP8B1 expression without altering the underlying DNA sequence.

CYP8B1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CYP8B1 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 CYP8B1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CYP8B1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CYP8B1 locus and enabling the study of CYP8B1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CYP8B1 pathway restoration in tumor cells with silenced or reduced CYP8B1 expression.

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