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.
Orai1 Double Nickase Plasmid (h): sc-400901-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Orai1 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
- Orai1 Double Nickase Plasmid (h) and Orai1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ORAI1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Orai1 Antibody (G-2): sc-377281
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Orai1 Double Nickase Plasmid (h) | sc-400901-NIC | 20 µg | $410.00 | |||
Orai1 Double Nickase Plasmid (h2) | sc-400901-NIC-2 | 20 µg | $410.00 |
ORAI1 encodes Orai1, the pore-forming subunit of the calcium release–activated calcium (CRAC) channel that mediates store-operated Ca²⁺ entry following depletion of endoplasmic reticulum Ca²⁺ stores. Orai1 cooperates with STIM1 to couple ER Ca²⁺ sensing to plasma membrane channel opening, shaping cytosolic calcium signals that regulate NFAT-dependent transcription, cell activation programs, and calcium homeostasis. This pathway is central to immune cell signaling, secretion, and broader calcium-regulated processes such as proliferation and migration. Dysregulated ORAI1 activity has been linked to immune dysfunction and inflammatory phenotypes, and altered Ca²⁺ influx signaling has been investigated in contexts including myopathy and cancer-associated signaling networks.
Orai1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ORAI1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ORAI1. 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 ORAI1 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 ORAI1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.