Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
RAGE Double Nickase Plasmid (h): sc-400284-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- RAGE 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
- RAGE Double Nickase Plasmid (h) and RAGE Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting AGER. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: RAGE Antibody (A-9): sc-365154
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
RAGE Double Nickase Plasmid (h) | sc-400284-NIC | 20 µg | $410.00 | |||
RAGE Double Nickase Plasmid (h2) | sc-400284-NIC-2 | 20 µg | $410.00 |
AGER encodes the receptor for advanced glycation end products (RAGE), a multiligand pattern-recognition receptor of the immunoglobulin superfamily that binds AGEs, S100 proteins, HMGB1, and other damage-associated molecules. RAGE signaling amplifies inflammatory and oxidative stress responses through pathways including NF-κB, MAPK/ERK, JNK/p38, PI3K/AKT, and Rho GTPase-dependent cytoskeletal remodeling. Sustained receptor engagement can promote cytokine production, adhesion molecule expression, and altered barrier function, linking RAGE biology to chronic inflammation and tissue remodeling. Dysregulated AGER/RAGE activity has been implicated in diabetes-associated vascular complications, atherosclerosis, neuroinflammation, and tumor-associated inflammation, making it a frequent target in mechanistic studies of innate immune signaling.
RAGE Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the AGER locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within AGER. 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 AGER 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 AGER-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.