BRAF Double Nickase Plasmid (h): sc-400121-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- BRAF 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
- BRAF Double Nickase Plasmid (h) and BRAF Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting BRAF. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: BRAF Antibody (F-7): sc-5284
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
BRAF Double Nickase Plasmid (h) | sc-400121-NIC | 20 µg | $410.00 | |||
BRAF Double Nickase Plasmid (h2) | sc-400121-NIC-2 | 20 µg | $410.00 |
BRAF encodes a serine/threonine protein kinase that functions as a key effector downstream of RAS to propagate signaling through the RAF–MEK–ERK/MAPK cascade. By regulating phosphorylation-dependent control of transcriptional programs, cell-cycle progression, differentiation, and survival, BRAF integrates extracellular cues into coordinated cellular responses. Dysregulated BRAF activity is linked to aberrant MAPK signaling and altered growth control in multiple disease-relevant contexts, making it a widely used node for dissecting oncogenic signaling networks. Human BRAF is therefore frequently interrogated in pathway mapping, genotype–phenotype studies, and analyses of signaling feedback and crosstalk with PI3K/AKT and stress-response pathways.
BRAF Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the BRAF locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within BRAF. 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 BRAF 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 BRAF-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.