NEDD4 Double Nickase Plasmid (h): sc-401748-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- NEDD4 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
- NEDD4 Double Nickase Plasmid (h) and NEDD4 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting NEDD4. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: NEDD4 Antibody (F-11): sc-518160
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
NEDD4 Double Nickase Plasmid (h) | sc-401748-NIC | 20 µg | $410.00 | |||
NEDD4 Double Nickase Plasmid (h2) | sc-401748-NIC-2 | 20 µg | $410.00 |
Human NEDD4 encodes an E3 ubiquitin-protein ligase that catalyzes ubiquitin transfer to substrates to regulate protein turnover, trafficking, and signaling amplitude. Through its C2 and WW domains, NEDD4 recognizes PY-motif–containing targets and influences pathways such as PI3K–AKT signaling, endocytosis, and membrane receptor homeostasis. NEDD4-dependent ubiquitination modulates epithelial polarity and cell migration programs, and it has been linked to regulation of tumor suppressors and growth factor receptors in multiple cellular contexts. Dysregulated NEDD4 activity and substrate selection have been associated with cancer-related signaling rewiring and altered immune and neuronal processes, making it a useful node for mechanistic studies of ubiquitin-driven regulation.
NEDD4 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the NEDD4 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within NEDD4. 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 NEDD4 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 NEDD4-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.