3β-HSD CRISPR Activation Plasmid (h): sc-400825-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- 3β-HSD CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- 3β-HSD CRISPR Activation Plasmid (h) consists of three plasmids at a 1:1:1 mass ratio: a plasmid encoding the deactivated Cas9 (dCas9) nuclease (D10A and N863A) fused to the transactivation domain VP64, and a blasticidin resistance gene; a plasmid encoding the MS2-p65-HSF1 fusion protein, and a hygromycin resistance gene; a plasmid encoding a target-specific 20 nt guide RNA fused to two MS2 RNA aptamers, and a puromycin resistance gene
- The resulting SAM complex binds to a site-specific region approximately 200-250 nt upstream of the transcriptional start site and provides robust recruitment of transcription factors for highly efficient gene activation
- gRNAs encoded by 3β-HSD CRISPR Activation Plasmid (h) and 3β-HSD CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the HSD3B1 transcriptional start site. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
3β-HSD CRISPR Activation Plasmid (h) | sc-400825-ACT | 20 µg | $397.00 |
HSD3B1 encodes 3β-hydroxysteroid dehydrogenase/Δ5→Δ4 isomerase (3β-HSD), a key microsomal enzyme that converts Δ5-3β-hydroxysteroids to Δ4-ketosteroids during steroidogenesis. This catalytic step is essential for the biosynthesis of progesterone, androgens, glucocorticoids, and mineralocorticoids, linking HSD3B1 activity to endocrine homeostasis and lipid-derived signaling. 3β-HSD function intersects with mitochondrial and ER steroid metabolic networks, including cholesterol utilization and downstream nuclear receptor signaling. Dysregulated HSD3B1 expression or activity has been associated with altered steroid hormone balance and is frequently investigated in hormone-dependent disease biology and metabolism-focused cellular phenotypes.
3β-HSD CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HSD3B1 expression without altering the underlying DNA sequence.
3β-HSD CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HSD3B1 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 HSD3B1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous 3β-HSD expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HSD3B1 locus and enabling the study of 3β-HSD-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of 3β-HSD pathway restoration in tumor cells with silenced or reduced HSD3B1 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.