Factor VIII Double Nickase Plasmid (h): sc-400373-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Factor VIII 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
- Factor VIII Double Nickase Plasmid (h) and Factor VIII Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting F8. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Factor VIII light chain Antibody (RFFVIII C/5): sc-59512
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Factor VIII Double Nickase Plasmid (h) | sc-400373-NIC | 20 µg | $410.00 | |||
Factor VIII Double Nickase Plasmid (h2) | sc-400373-NIC-2 | 20 µg | $410.00 |
Human F8 encodes coagulation factor VIII, a glycoprotein cofactor that circulates bound to von Willebrand factor and is activated by thrombin during the intrinsic pathway. Activated factor VIII (FVIIIa) forms the intrinsic tenase complex with factor IXa on phospholipid surfaces, accelerating factor X activation and downstream thrombin generation within the coagulation cascade. Loss-of-function variants in F8 reduce intrinsic pathway efficiency and are strongly associated with hemophilia A, providing a molecular entry point for studying hemostasis, endothelial–hepatic protein biosynthesis, and protease cofactor regulation. Experimental perturbation of F8 supports mechanistic analysis of coagulation network dynamics, FVIII processing and secretion, and interactions with vWF in human cellular models.
Factor VIII Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the F8 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within F8. 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 F8 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 F8-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.