MDHC Double Nickase Plasmid (h): sc-416827-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- MDHC 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
- MDHC Double Nickase Plasmid (h) and MDHC Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting MDH1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: MDHC Antibody (H-6): sc-166879
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
MDHC Double Nickase Plasmid (h) | sc-416827-NIC | 20 µg | $410.00 | |||
MDHC Double Nickase Plasmid (h2) | sc-416827-NIC-2 | 20 µg | $410.00 |
MDH1 encodes the cytosolic malate dehydrogenase (MDHC), a NAD+/NADH-dependent enzyme that interconverts malate and oxaloacetate, linking glycolysis-derived carbon flux to the tricarboxylic acid cycle through the malate–aspartate shuttle. By supporting cytosolic redox balance and NADH transfer into mitochondria, MDHC helps coordinate energy metabolism, anaplerosis, and biosynthetic precursor availability. MDH1 activity is tightly connected to cellular responses to nutrient state and oxidative stress through regulation of cytosolic NADH/NAD+ ratios. Altered MDH1 expression or metabolic rewiring involving malate/oxaloacetate cycling has been observed in metabolic and proliferative disease contexts, motivating mechanistic studies in human cell models.
MDHC Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the MDH1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within MDH1. 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 MDH1 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 MDH1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.