SEMA5A Double Nickase Plasmid (m): sc-422885-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- SEMA5A Double Nickase Plasmid (m) 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
- SEMA5A Double Nickase Plasmid (m) and SEMA5A Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Sema5a. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
SEMA5A Double Nickase Plasmid (m) | sc-422885-NIC | 20 µg | $410.00 | |||
SEMA5A Double Nickase Plasmid (m2) | sc-422885-NIC-2 | 20 µg | $410.00 |
Mouse Sema5a encodes the transmembrane guidance cue SEMA5A, a member of the semaphorin family that modulates axon navigation, dendritic patterning, and cell motility through receptor interactions that influence cytoskeletal remodeling and adhesion dynamics. SEMA5A signaling interfaces with pathways controlling growth cone guidance and cell–cell/ECM interactions, contributing to neural circuit formation and tissue organization. Altered Sema5a expression or function has been associated with neurodevelopmental and neurobehavioral phenotypes, and dysregulated semaphorin signaling is also studied in contexts of aberrant migration and invasion programs. These features make Sema5a a useful target for dissecting mechanisms of developmental wiring, synaptic connectivity, and semaphorin-mediated regulation of cellular movement.
SEMA5A Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Sema5a locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Sema5a. 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 Sema5a 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 Sema5a-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.