α-defensin 5 Double Nickase Plasmid (h): sc-404991-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- α-defensin 5 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
- α-defensin 5 Double Nickase Plasmid (h) and α-defensin 5 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting DEFA5. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: α-defensin 5 Antibody (8c8): sc-53997
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
α-defensin 5 Double Nickase Plasmid (h) | sc-404991-NIC | 20 µg | $410.00 | |||
α-defensin 5 Double Nickase Plasmid (h2) | sc-404991-NIC-2 | 20 µg | $410.00 |
DEFA5 encodes human α-defensin 5, a cationic antimicrobial peptide produced predominantly by Paneth cells in the small intestine and released into the intestinal lumen as part of innate mucosal immunity. After proteolytic processing, α-defensin 5 disrupts microbial membranes and shapes the composition of the gut microbiota while supporting epithelial barrier function and host defense programs. Its expression is linked to Paneth cell differentiation and secretory granule biology, and it interfaces with inflammatory signaling and microbial sensing pathways that regulate intestinal homeostasis. Altered DEFA5 levels and Paneth cell dysfunction have been associated with inflammatory bowel conditions and susceptibility to enteric infection, making it a useful marker and functional node in mucosal immunology research.
α-defensin 5 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the DEFA5 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within DEFA5. 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 DEFA5 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 DEFA5-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.