Thrombin R Double Nickase Plasmid (h): sc-400556-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Thrombin R 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
- Thrombin R Double Nickase Plasmid (h) and Thrombin R Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting F2R. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Thrombin R Antibody (ATAP2): sc-13503
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Thrombin R Double Nickase Plasmid (h) | sc-400556-NIC | 20 µg | $410.00 | |||
Thrombin R Double Nickase Plasmid (h2) | sc-400556-NIC-2 | 20 µg | $410.00 |
F2R encodes thrombin receptor (protease-activated receptor-1, PAR1), a GPCR activated by thrombin-mediated proteolytic cleavage that exposes a tethered ligand and initiates signaling through Gαq, Gα12/13, and Gαi pathways. PAR1 activation regulates platelet activation, endothelial barrier function, vascular tone, and leukocyte trafficking via downstream MAPK, RhoA/ROCK, PI3K-AKT, and NF-κB signaling. Through integration of coagulation and inflammatory cues, F2R contributes to hemostasis, thromboinflammation, and tissue remodeling. Dysregulated PAR1 signaling has been associated with thrombosis, atherosclerosis, sepsis-associated coagulopathy, fibrosis, and tumor microenvironment interactions, supporting mechanistic studies across vascular biology and cancer research contexts.
Thrombin R Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the F2R locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within F2R. 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 F2R 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 F2R-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.