CYP11B1 CRISPR Activation Plasmid (h): sc-403040-ACT

CYP11B1 encodes cytochrome P450 11β-hydroxylase, a mitochondrial monooxygenase that catalyzes late steps in adrenal steroidogenesis, including the conversion of 11-deoxycortisol to cortisol and 11-deoxycorticosterone to corticosterone. This enzyme functions within heme-dependent redox reactions requiring electron transfer from adrenodoxin/adrenodoxin reductase, linking mitochondrial metabolism to hormonal output. CYP11B1 activity influences hypothalamic–pituitary–adrenal axis regulation and downstream transcriptional programs responsive to glucocorticoids and mineralocorticoids. Altered CYP11B1 expression or function is associated with congenital adrenal hyperplasia phenotypes and contributes to endocrine dysregulation that is relevant for modeling adrenal disorders and steroid-driven disease states.

CYP11B1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CYP11B1 expression without altering the underlying DNA sequence.

CYP11B1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CYP11B1 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 CYP11B1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CYP11B1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CYP11B1 locus and enabling the study of CYP11B1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CYP11B1 pathway restoration in tumor cells with silenced or reduced CYP11B1 expression.

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