A-Raf Double Nickase Plasmid (h): sc-401799-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- A-Raf 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
- A-Raf Double Nickase Plasmid (h) and A-Raf Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ARAF. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: A-Raf Antibody (A-5): sc-166771
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
A-Raf Double Nickase Plasmid (h) | sc-401799-NIC | 20 µg | $410.00 | |||
A-Raf Double Nickase Plasmid (h2) | sc-401799-NIC-2 | 20 µg | $410.00 |
ARAF encodes the serine/threonine kinase A‑Raf, a RAF family member that links activated RAS to the MAPK/ERK signaling cascade. Through regulated activation and dimerization, A‑Raf contributes to phosphorylation of MEK and downstream control of cell proliferation, differentiation, and survival programs. ARAF activity intersects with growth factor receptor signaling and can influence feedback regulation within the ERK pathway and cross-talk with PI3K–AKT signaling. Dysregulated RAF–MEK–ERK signaling, including altered RAF isoform usage, is broadly relevant to oncogenic signaling networks and to modeling context-dependent pathway dependencies in human cells.
A-Raf Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ARAF locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ARAF. 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 ARAF 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 ARAF-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.