EphA3 Double Nickase Plasmid (h): sc-401565-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- EphA3 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
- EphA3 Double Nickase Plasmid (h) and EphA3 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting EPHA3. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: EphA3 Antibody (D-2): sc-514209
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
EphA3 Double Nickase Plasmid (h) | sc-401565-NIC | 20 µg | $410.00 | |||
EphA3 Double Nickase Plasmid (h2) | sc-401565-NIC-2 | 20 µg | $410.00 |
EPHA3 encodes EphA3, a receptor tyrosine kinase in the ephrin-A signaling network that regulates cell–cell communication, tissue patterning, and cytoskeletal remodeling. Upon ephrin engagement, EphA3 influences bidirectional signaling that intersects with Rho-family GTPases, MAPK, and PI3K-associated pathways to modulate cell adhesion, migration, and boundary formation. Dysregulated EPHA3 expression or signaling has been linked to altered developmental programs and aberrant growth and invasion phenotypes across multiple cancer contexts, making it a useful node for studying receptor-driven guidance and tumor microenvironment interactions. In human cell models, EphA3-dependent signaling provides a tractable system to interrogate contact-dependent signaling dynamics and downstream transcriptional responses.
EphA3 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the EPHA3 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within EPHA3. 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 EPHA3 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 EPHA3-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.