PAR-4 Double Nickase Plasmid (h): sc-401719-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PAR-4 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
- PAR-4 Double Nickase Plasmid (h) and PAR-4 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting F2RL3. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: PAR-4 Antibody (5F4): sc-293206
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
PAR-4 Double Nickase Plasmid (h) | sc-401719-NIC | 20 µg | $410.00 | |||
PAR-4 Double Nickase Plasmid (h2) | sc-401719-NIC-2 | 20 µg | $410.00 |
F2RL3 encodes protease-activated receptor-4 (PAR-4), a thrombin-responsive GPCR that is activated by proteolytic cleavage to unmask a tethered ligand, initiating signaling through Gq/11 and G12/13. PAR-4 engagement promotes phospholipase C–dependent calcium mobilization, PKC activation, and RhoA-mediated cytoskeletal remodeling, integrating with MAPK/ERK pathways to regulate platelet activation, granule secretion, and thrombus stabilization. Beyond hemostasis, F2RL3/PAR-4 activity contributes to inflammatory signaling in vascular and immune contexts and has been investigated in cardiometabolic and vascular disease biology, including atherosclerosis-related processes. Its expression and signaling dynamics also provide a mechanistic entry point for studying protease–GPCR crosstalk and coagulation-linked inflammation in relevant human cell models.
PAR-4 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the F2RL3 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within F2RL3. 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 F2RL3 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 F2RL3-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.