DLD Double Nickase Plasmid (h): sc-403207-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- DLD 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
- DLD Double Nickase Plasmid (h) and DLD Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting DLD. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: DLD Antibody (G-2): sc-365977
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
DLD Double Nickase Plasmid (h) | sc-403207-NIC | 20 µg | $410.00 | |||
DLD Double Nickase Plasmid (h2) | sc-403207-NIC-2 | 20 µg | $410.00 |
DLD encodes dihydrolipoamide dehydrogenase (E3), a flavin-dependent oxidoreductase that reoxidizes dihydrolipoamide to lipoamide while reducing NAD+ to NADH. As a shared E3 subunit, DLD supports mitochondrial energy metabolism through the pyruvate dehydrogenase, α-ketoglutarate dehydrogenase, and branched-chain α-ketoacid dehydrogenase complexes, linking glycolysis and amino acid catabolism to the TCA cycle and oxidative phosphorylation. Disruption of DLD perturbs redox homeostasis and mitochondrial function, altering NADH/NAD+ balance and reactive oxygen species handling. Genetic variation in DLD is associated with inborn errors of metabolism characterized by impaired oxidative metabolism and lactic acidemia, and dysregulated mitochondrial energetics is relevant to neurodegeneration and cancer bioenergetics research.
DLD Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the DLD locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within DLD. 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 DLD 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 DLD-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.