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.
CYP27B1 Double Nickase Plasmid (h): sc-401155-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- CYP27B1 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
- CYP27B1 Double Nickase Plasmid (h) and CYP27B1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting CYP27B1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: CYP27B1 Antibody (G-5): sc-515903
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
CYP27B1 Double Nickase Plasmid (h) | sc-401155-NIC | 20 µg | $410.00 | |||
CYP27B1 Double Nickase Plasmid (h2) | sc-401155-NIC-2 | 20 µg | $410.00 |
CYP27B1 encodes mitochondrial 25-hydroxyvitamin D-1α-hydroxylase, the key enzyme that converts 25-hydroxyvitamin D to the active hormone 1,25-dihydroxyvitamin D (calcitriol). By controlling calcitriol abundance, CYP27B1 regulates vitamin D receptor (VDR) signaling programs that influence calcium and phosphate homeostasis, bone mineralization, and broad transcriptional networks in immune and epithelial cells. Its activity links sterol metabolism to endocrine signaling and mitochondrial redox processes, with downstream effects on differentiation, inflammatory tone, and barrier biology. Dysregulated CYP27B1 expression or function is associated with disorders of vitamin D metabolism and has been studied in contexts including skeletal phenotypes, autoimmune susceptibility, chronic inflammation, and altered tumor microenvironment signaling.
CYP27B1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the CYP27B1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within CYP27B1. 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 CYP27B1 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 CYP27B1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.