Raf-1 Double Nickase Plasmid (h): sc-400202-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Raf-1 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
- Raf-1 Double Nickase Plasmid (h) and Raf-1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting RAF1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Raf-1 Antibody (E-10): sc-7267
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Raf-1 Double Nickase Plasmid (h) | sc-400202-NIC | 20 µg | $410.00 | |||
Raf-1 Double Nickase Plasmid (h2) | sc-400202-NIC-2 | 20 µg | $410.00 |
RAF1 encodes the serine/threonine kinase Raf-1, a central effector of RAS signaling that relays inputs from activated receptor tyrosine kinases to the MAPK/ERK cascade. Raf-1 regulates cell-cycle progression, differentiation, apoptosis, and stress responses through phosphorylation of downstream kinases including MEK1/2 and consequent ERK1/2 activation. Dysregulated RAF1 activity and aberrant MAPK pathway signaling are implicated in oncogenic transformation and resistance mechanisms, and germline or somatic alterations in RAF1 have been linked to developmental syndromes and cardiomyopathy-associated signaling defects. As a signaling hub that integrates growth factor and inflammatory cues, Raf-1 is widely studied in pathway crosstalk with PI3K-AKT, feedback regulation, and phosphatase-mediated control of kinase activity.
Raf-1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the RAF1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within RAF1. 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 RAF1 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 RAF1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.