NEDL2 Double Nickase Plasmid (m): sc-435931-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- NEDL2 Double Nickase Plasmid (m) 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
- NEDL2 Double Nickase Plasmid (m) and NEDL2 Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Hecw2. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
NEDL2 Double Nickase Plasmid (m) | sc-435931-NIC | 20 µg | $410.00 | |||
NEDL2 Double Nickase Plasmid (m2) | sc-435931-NIC-2 | 20 µg | $410.00 |
Hecw2 encodes NEDL2, a HECT-type E3 ubiquitin ligase that mediates substrate ubiquitination to control protein stability and signaling output. NEDL2 activity contributes to proteostasis and regulation of cellular processes such as neuronal development, synaptic function, and stress-responsive pathways through targeted degradation of regulatory proteins. In mouse models and human genetics, HECW2/NEDL2 has been linked to neurodevelopmental phenotypes, including altered neuronal maturation and signaling defects, making it relevant for mechanistic studies of nervous system biology. As an E3 ligase, NEDL2 provides a node to interrogate ubiquitin-dependent control of pathway dynamics and cell state transitions.
NEDL2 Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Hecw2 locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Hecw2. 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 Hecw2 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 Hecw2-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.