nestin Double Nickase Plasmid (h): sc-400156-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- nestin 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
- nestin Double Nickase Plasmid (h) and nestin Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting NES. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: nestin Antibody (D-9): sc-377380
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
nestin Double Nickase Plasmid (h) | sc-400156-NIC | 20 µg | $410.00 | |||
nestin Double Nickase Plasmid (h2) | sc-400156-NIC-2 | 20 µg | $410.00 |
NES encodes nestin, a class VI intermediate filament protein enriched in neural stem and progenitor cells where it supports cytoskeletal organization during proliferation, migration, and lineage commitment. Nestin participates in cytoskeletal remodeling and cell cycle–linked programs, integrating cues from developmental signaling pathways that regulate stemness and differentiation, including Notch, Wnt/β-catenin, and growth factor–driven MAPK/PI3K-AKT signaling. Its expression is widely used as a marker of neuroepithelial progenitors and reactive astroglia, and it is frequently induced during tissue remodeling and cellular stress responses. Dysregulated NES expression is associated with altered differentiation states and has been reported in multiple cancers and neurodevelopmental contexts, making it relevant for studying progenitor biology and disease-associated cell state transitions.
nestin Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the NES locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within NES. 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 NES 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 NES-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.