EphA5 Double Nickase Plasmid (h): sc-403104-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- EphA5 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
- EphA5 Double Nickase Plasmid (h) and EphA5 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting EPHA5. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: EphA5 Antibody (8B10B1): sc-517242
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
EphA5 Double Nickase Plasmid (h) | sc-403104-NIC | 20 µg | $410.00 | |||
EphA5 Double Nickase Plasmid (h2) | sc-403104-NIC-2 | 20 µg | $410.00 |
EPHA5 encodes EphA5, a receptor tyrosine kinase in the ephrin-A signaling system that governs contact-dependent communication between neighboring cells. EphA5 modulates axon guidance, synapse formation, and neuronal circuit refinement through bidirectional signaling that impacts cytoskeletal remodeling, adhesion, and endocytic trafficking. Downstream pathways commonly linked to Eph receptors include Rho family GTPases, MAPK/ERK, and PI3K-related signaling nodes that integrate cues for migration and neurite dynamics. Altered EPHA5 regulation and ephrin pathway imbalance have been associated with neurodevelopmental phenotypes and context-dependent roles in oncogenic processes such as invasion and cell–cell boundary formation, supporting mechanistic studies in both neural and cancer models.
EphA5 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the EPHA5 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within EPHA5. 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 EPHA5 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 EPHA5-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.