Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Prothrombin Double Nickase Plasmid (h): sc-400962-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Prothrombin 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
- Prothrombin Double Nickase Plasmid (h) and Prothrombin Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting F2. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Thrombin Antibody (F-1): sc-271449
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Prothrombin Double Nickase Plasmid (h) | sc-400962-NIC | 20 µg | $410.00 | |||
Prothrombin Double Nickase Plasmid (h2) | sc-400962-NIC-2 | 20 µg | $410.00 |
F2 encodes human prothrombin, a vitamin K–dependent zymogen that is proteolytically converted to thrombin to drive the central steps of the coagulation cascade. Thrombin cleaves fibrinogen to fibrin, amplifies clot formation via activation of factors V, VIII, XI, and XIII, and signals through protease-activated receptors to coordinate platelet activation and vascular inflammatory responses. Prothrombin production and maturation depend on hepatic secretory processing and γ-carboxylation, linking F2 regulation to nutrient status and endoplasmic reticulum quality control. Dysregulated F2 activity and genetic variation are widely studied in the context of thrombosis and hemostatic imbalance, providing a mechanistic entry point for dissecting coagulation pathway dynamics in cellular and in vivo models.
Prothrombin Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the F2 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within F2. 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 F2 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 F2-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.