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.
PDH-E1α Double Nickase Plasmid (h): sc-401044-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PDH-E1α 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
- PDH-E1α Double Nickase Plasmid (h) and PDH-E1α Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting PDHA1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: PDH-E1α Antibody (D-6): sc-377092
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
PDH-E1α Double Nickase Plasmid (h) | sc-401044-NIC | 20 µg | $410.00 | |||
PDH-E1α Double Nickase Plasmid (h2) | sc-401044-NIC-2 | 20 µg | $410.00 |
PDHA1 encodes the E1α subunit of the pyruvate dehydrogenase (PDH) complex, which catalyzes the oxidative decarboxylation of pyruvate to generate acetyl-CoA and link glycolysis to the tricarboxylic acid cycle. PDH-E1α activity supports mitochondrial respiration, regulates carbon flux into lipid biosynthesis, and influences redox balance through downstream NADH production. PDHA1 is regulated by PDH kinases and phosphatases that tune PDH complex activity in response to nutrient availability and cellular energy demand. Perturbation of PDHA1 function is associated with metabolic remodeling and mitochondrial dysfunction, and is studied in contexts such as lactic acidosis, neurodevelopmental phenotypes, and altered bioenergetics in proliferative disease models.
PDH-E1α Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the PDHA1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within PDHA1. 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 PDHA1 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 PDHA1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.