CD100 Double Nickase Plasmid (h): sc-405956-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- CD100 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
- CD100 Double Nickase Plasmid (h) and CD100 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting SEMA4D. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: CD100 Antibody (C-3): sc-390675
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
CD100 Double Nickase Plasmid (h) | sc-405956-NIC | 20 µg | $410.00 | |||
CD100 Double Nickase Plasmid (h2) | sc-405956-NIC-2 | 20 µg | $410.00 |
SEMA4D encodes CD100, a transmembrane semaphorin that also exists as a soluble ectodomain and functions in bidirectional signaling through receptors such as plexin-B1/B2 and CD72. CD100 regulates immune cell activation, B cell signaling thresholds, cytokine production, and leukocyte migration, and it also influences neuronal guidance and angiogenic responses via Rho-family GTPase and PI3K-associated pathways. Through these processes, CD100 is implicated in shaping tumor–immune interactions, inflammatory tissue remodeling, and neuroinflammatory mechanisms. Dysregulated SEMA4D/CD100 signaling has been associated with altered immune surveillance and pathological inflammation across multiple disease contexts.
CD100 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the SEMA4D locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within SEMA4D. 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 SEMA4D 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 SEMA4D-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.