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.
IDH2 Double Nickase Plasmid (h): sc-401417-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- IDH2 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
- IDH2 Double Nickase Plasmid (h) and IDH2 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting IDH2. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: IDH2 Antibody (B-6): sc-374476
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
IDH2 Double Nickase Plasmid (h) | sc-401417-NIC | 20 µg | $410.00 | |||
IDH2 Double Nickase Plasmid (h2) | sc-401417-NIC-2 | 20 µg | $410.00 |
IDH2 encodes the mitochondrial NADP-dependent isocitrate dehydrogenase that catalyzes oxidative decarboxylation of isocitrate to α-ketoglutarate while producing NADPH. This reaction supports the tricarboxylic acid cycle, mitochondrial redox homeostasis, and antioxidant defenses by maintaining NADPH-dependent glutathione and thioredoxin pathways. IDH2 activity influences cellular differentiation programs and metabolic adaptation under oxidative stress, and dysregulation of IDH2 function is linked to altered α-ketoglutarate metabolism and epigenetic regulation. Recurrent IDH2 mutations that generate the oncometabolite 2-hydroxyglutarate are associated with aberrant DNA and histone methylation patterns, making IDH2 a key node in studies of cancer metabolism and mitochondrial signaling.
IDH2 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the IDH2 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within IDH2. 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 IDH2 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 IDH2-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.