Cryopyrin/NALP3/NLRP3 Double Nickase Plasmid (h): sc-400270-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Cryopyrin/NALP3/NLRP3 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
- Cryopyrin/NALP3/NLRP3 Double Nickase Plasmid (h) and Cryopyrin/NALP3/NLRP3 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting NLRP3. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Cryopyrin/NALP3/NLRP3 Antibody (6F12): sc-134306
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Cryopyrin/NALP3/NLRP3 Double Nickase Plasmid (h) | sc-400270-NIC | 20 µg | $410.00 | |||
Cryopyrin/NALP3/NLRP3 Double Nickase Plasmid (h2) | sc-400270-NIC-2 | 20 µg | $410.00 |
Human NLRP3 (cryopyrin/NALP3) is a cytosolic pattern-recognition receptor that nucleates the NLRP3 inflammasome in response to diverse pathogen- and danger-associated cues, including ionic flux, mitochondrial stress, and lysosomal damage. Upon activation, NLRP3 assembles with ASC and pro-caspase-1 to drive caspase-1 activation, promoting maturation of IL-1β and IL-18 and triggering gasdermin D–dependent pyroptosis. This signaling axis is central to innate immune regulation and links metabolic and inflammatory stress to downstream cytokine programs. Dysregulated NLRP3 activity and variants are associated with autoinflammatory syndromes and contribute to inflammatory mechanisms studied in gout, atherosclerosis, neuroinflammation, and metabolic disease models.
Cryopyrin/NALP3/NLRP3 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the NLRP3 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within NLRP3. 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 NLRP3 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 NLRP3-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.