CYP17A1 CRISPR Activation Plasmid (h): sc-401918-ACT

CYP17A1 encodes cytochrome P450 17A1, a microsomal monooxygenase that catalyzes both 17α-hydroxylase and 17,20-lyase reactions in steroidogenic tissues, directing flux through glucocorticoid and sex steroid biosynthesis. As a key node in the steroid hormone pathway, CYP17A1 influences downstream androgen and estrogen production and contributes to endocrine regulation through coordinated activity with other P450 enzymes and redox partners in the endoplasmic reticulum. Altered CYP17A1 expression or enzymatic function has been associated with steroidogenic imbalance and endocrine phenotypes, making it a useful target for mechanistic studies of hormone metabolism and signal-dependent transcriptional programs. Experimental modulation of CYP17A1 supports investigations into steroid-driven cell states, metabolic rewiring, and pathway crosstalk relevant to endocrine biology.

CYP17A1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CYP17A1 expression without altering the underlying DNA sequence.

CYP17A1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CYP17A1 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 CYP17A1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CYP17A1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CYP17A1 locus and enabling the study of CYP17A1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CYP17A1 pathway restoration in tumor cells with silenced or reduced CYP17A1 expression.

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