Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
3β-HSD Double Nickase Plasmid (h): sc-400825-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- 3β-HSD Double Nickase Plasmid (h) consists of a pair of plasmids each encoding a D10A mutated Cas9 nuclease and a target-specific 20 nt guide RNA (gRNA) designed to knockout gene expression with greater specificity than its CRISPR/Cas9 KO counterpart
- Paired gRNA sequences are offset by approximately 20 bp to allow for specific Cas9-mediated double nicking of the genomic DNA, which mimics a DSB
- One plasmid in the pair contains a puromycin-resistance gene for selection; the other plasmid in the pair contains a GFP marker to visually confirm transfection
- 3β-HSD Double Nickase Plasmid (h) and 3β-HSD Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting HSD3B1. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
3β-HSD Double Nickase Plasmid (h) | sc-400825-NIC | 20 µg | $410.00 | |||
3β-HSD Double Nickase Plasmid (h2) | sc-400825-NIC-2 | 20 µg | $410.00 |
HSD3B1 encodes 3β-hydroxysteroid dehydrogenase/Δ5-Δ4 isomerase type 1 (3β-HSD), a microsomal enzyme that catalyzes key steps in steroidogenesis by converting Δ5-3β-hydroxysteroids to their Δ4-ketosteroid products, including the conversion of pregnenolone to progesterone and dehydroepiandrosterone to androstenedione. Through these reactions it regulates flux through androgen, estrogen, glucocorticoid, and mineralocorticoid biosynthetic pathways and influences cellular steroid hormone availability. Altered HSD3B1 activity or expression can shift intracrine steroid metabolism and is studied in contexts where hormone-dependent signaling and differentiation are perturbed, including endocrine and reproductive biology and hormone-responsive disease models.
3β-HSD Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the HSD3B1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within HSD3B1. When directed to adjacent sites on opposite DNA strands, the two nickases generate offset single-strand nicks that together produce a staggered double-strand break, requiring coordinated on-target activity from both guides. The resulting DNA break is resolved by endogenous cellular repair pathways, most commonly through non-homologous end joining (NHEJ), leading to insertions or deletions that disrupt HSD3B1 function. By requiring dual sgRNA engagement at the target locus, the double nicking approach enhances editing specificity and provides a complementary CRISPR strategy for applications where additional control over targeting precision is desired.
To support efficient identification of edited cells, one plasmid encodes GFP for fluorescent visualization of transfected populations, while the companion plasmid carries a puromycin resistance gene for antibiotic selection. Together, these features support efficient enrichment of co-transfected populations and simplify the validation of HSD3B1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.