Neurofibromin Double Nickase Plasmid (h): sc-400625-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Neurofibromin 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
- Neurofibromin Double Nickase Plasmid (h) and Neurofibromin Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting NF1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Neurofibromin Antibody (H-12): sc-376886
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Neurofibromin Double Nickase Plasmid (h) | sc-400625-NIC | 20 µg | $410.00 | |||
Neurofibromin Double Nickase Plasmid (h2) | sc-400625-NIC-2 | 20 µg | $410.00 |
NF1 encodes neurofibromin, a large cytoplasmic tumor suppressor that functions primarily as a Ras GTPase-activating protein (Ras-GAP), converting active Ras-GTP to Ras-GDP to restrain downstream MAPK/ERK and PI3K-AKT signaling. Through this negative regulation, neurofibromin influences cell-cycle progression, differentiation, and survival, and contributes to homeostatic control of growth factor signaling. Loss-of-function alterations in NF1 disrupt Ras pathway regulation and are strongly associated with neurofibromatosis type 1 and with Ras-driven oncogenic processes in multiple tissue contexts. NF1 perturbation is therefore widely used to study signal transduction, cellular plasticity, and genotype–phenotype relationships in disease-relevant models.
Neurofibromin Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the NF1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within NF1. 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 NF1 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 NF1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.