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.
EphB4 Double Nickase Plasmid (h): sc-401122-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- EphB4 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
- EphB4 Double Nickase Plasmid (h) and EphB4 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting EPHB4. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: EphB4 Antibody (H-10): sc-365510
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
EphB4 Double Nickase Plasmid (h) | sc-401122-NIC | 20 µg | $410.00 | |||
EphB4 Double Nickase Plasmid (h2) | sc-401122-NIC-2 | 20 µg | $410.00 |
EPHB4 encodes the EphB4 receptor tyrosine kinase, a key mediator of contact-dependent signaling through ephrin-B ligands that coordinates cell positioning, boundary formation, and vascular morphogenesis. Upon ephrin engagement, EphB4 triggers bidirectional signaling that intersects with Rho family GTPases, Src/FAK, PI3K–AKT, and MAPK pathways to regulate cytoskeletal dynamics, adhesion, and migration. EphB4 activity is strongly linked to endothelial biology and arteriovenous specification, and dysregulation of EPHB4–ephrin signaling has been associated with altered angiogenic behavior and invasive phenotypes in multiple disease contexts, including cancer and vascular malformations. These properties make EPHB4 a widely used target for mechanistic studies of cell–cell communication, vessel development, and microenvironmental signaling.
EphB4 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the EPHB4 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within EPHB4. 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 EPHB4 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 EPHB4-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.