CYP21A2 Double Nickase Plasmid (h): sc-402919-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- CYP21A2 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
- CYP21A2 Double Nickase Plasmid (h) and CYP21A2 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting CYP21A2. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: CYP21A2 Antibody (B-7): sc-518265
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
CYP21A2 Double Nickase Plasmid (h) | sc-402919-NIC | 20 µg | $410.00 | |||
CYP21A2 Double Nickase Plasmid (h2) | sc-402919-NIC-2 | 20 µg | $410.00 |
CYP21A2 encodes the cytochrome P450 enzyme steroid 21-hydroxylase, an endoplasmic reticulum–associated monooxygenase required for adrenal steroidogenesis. It catalyzes key hydroxylation steps in the biosynthetic pathways leading to cortisol and aldosterone, thereby influencing glucocorticoid and mineralocorticoid homeostasis. CYP21A2 activity integrates with mitochondrial and microsomal steroidogenic networks, including electron transfer from NADPH via P450 oxidoreductase and coordinated regulation of adrenal zonation programs. Loss-of-function variants in CYP21A2 are strongly associated with congenital adrenal hyperplasia due to 21-hydroxylase deficiency, making it a central target for studying steroid biosynthesis and endocrine disease mechanisms in human cell models.
CYP21A2 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the CYP21A2 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within CYP21A2. 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 CYP21A2 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 CYP21A2-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.