EDG-1/S1P1/S1PR1 Double Nickase Plasmid (h): sc-400413-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- EDG-1/S1P1/S1PR1 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
- EDG-1/S1P1/S1PR1 Double Nickase Plasmid (h) and EDG-1/S1P1/S1PR1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting S1PR1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: EDG-1/S1P1/S1PR1 Antibody (A-6): sc-48356
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
EDG-1/S1P1/S1PR1 Double Nickase Plasmid (h) | sc-400413-NIC | 20 µg | $410.00 | |||
EDG-1/S1P1/S1PR1 Double Nickase Plasmid (h2) | sc-400413-NIC-2 | 20 µg | $410.00 |
S1PR1 (EDG-1/S1P1) encodes a G protein–coupled receptor for sphingosine-1-phosphate that regulates endothelial barrier integrity, vascular maturation, and immune cell trafficking. Receptor engagement activates Gi-dependent signaling with downstream PI3K–AKT, MAPK/ERK, and Rho family GTPase pathways that shape cytoskeletal dynamics, adherens junction organization, and chemotactic responses. In hematopoietic and vascular contexts, S1PR1 contributes to lymphocyte egress, angiogenic sprouting, and inflammatory tone through integration with cytokine and chemokine networks. Dysregulated S1PR1 signaling has been associated with altered vascular permeability and immune homeostasis in inflammation, cardiovascular biology, and tumor microenvironment studies, supporting its use as a mechanistic node in receptor signaling research.
EDG-1/S1P1/S1PR1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the S1PR1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within S1PR1. 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 S1PR1 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 S1PR1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.