Pyrin Double Nickase Plasmid (h): sc-403193-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Pyrin 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
- Pyrin Double Nickase Plasmid (h) and Pyrin Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting MEFV. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Pyrin Antibody (C-11): sc-390938
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Pyrin Double Nickase Plasmid (h) | sc-403193-NIC | 20 µg | $410.00 | |||
Pyrin Double Nickase Plasmid (h2) | sc-403193-NIC-2 | 20 µg | $410.00 |
MEFV encodes pyrin, a myeloid-enriched innate immune sensor that regulates inflammasome signaling and inflammatory caspase activation in response to cytoskeletal and Rho GTPase–modulating cues. Pyrin helps coordinate ASC-dependent assembly of inflammasome complexes, influencing IL-1 family cytokine maturation and pyroptotic pathways that shape neutrophil-driven inflammation. Dysregulated MEFV activity or variants are associated with autoinflammatory phenotypes and altered thresholding of innate immune responses, making it a useful locus for dissecting inflammatory signaling networks. In human cell systems, MEFV perturbation is commonly used to study inflammasome regulation, cytokine processing, and crosstalk between cytoskeletal dynamics and innate immune sensing.
Pyrin Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the MEFV locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within MEFV. 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 MEFV 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 MEFV-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.