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.
GAPDH Double Nickase Plasmid (h): sc-400002-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- GAPDH 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
- GAPDH Double Nickase Plasmid (h) and GAPDH Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting GAPDH. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: GAPDH Antibody (0411): sc-47724
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
GAPDH Double Nickase Plasmid (h) | sc-400002-NIC | 20 µg | $410.00 | |||
GAPDH Double Nickase Plasmid (h2) | sc-400002-NIC-2 | 20 µg | $410.00 |
GAPDH encodes glyceraldehyde-3-phosphate dehydrogenase, a conserved cytosolic enzyme that catalyzes a key step in glycolysis linking glucose metabolism to ATP production and redox homeostasis via NAD+/NADH cycling. Beyond its metabolic function, GAPDH participates in cellular stress responses, including regulation of apoptosis, RNA binding, and nuclear signaling under oxidative or nitrosative conditions. As a central node in energy metabolism, GAPDH expression and activity are frequently modulated during hypoxia and metabolic rewiring, processes relevant to tumor biology, neurodegeneration, and inflammatory states. Its ubiquitous, high baseline expression also makes GAPDH a common reference target in gene expression and proteomics workflows, where perturbation can help assess normalization bias and metabolic confounders.
GAPDH Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the GAPDH locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within GAPDH. 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 GAPDH 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 GAPDH-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.