DFNA5 Double Nickase Plasmid (h): sc-410760-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- DFNA5 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
- DFNA5 Double Nickase Plasmid (h) and DFNA5 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting DFNA5. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: DFNA5 Antibody (G-9): sc-393162
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
DFNA5 Double Nickase Plasmid (h) | sc-410760-NIC | 20 µg | $410.00 |
DFNA5 (also known as GSDME) encodes a gasdermin family protein that links protease activation to membrane pore formation and programmed cell death. Upon caspase-3 cleavage, DFNA5 can drive a lytic, inflammatory cell-death program that intersects apoptosis-to-pyroptosis switching and influences cytokine release, membrane integrity, and innate immune signaling. DFNA5 activity has been implicated in epithelial stress responses and tumor biology, and germline variants are associated with autosomal dominant nonsyndromic hearing loss (DFNA5). These features make DFNA5 a useful node for studying regulated cell death, inflammatory signaling, and tissue-specific vulnerability mechanisms.
DFNA5 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the DFNA5 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within DFNA5. 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 DFNA5 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 DFNA5-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.