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.
GPT Double Nickase Plasmid (h): sc-402909-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- GPT 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
- GPT Double Nickase Plasmid (h) and GPT Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting GPT. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: GPT Antibody (E-3): sc-374501
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
GPT Double Nickase Plasmid (h) | sc-402909-NIC | 20 µg | $410.00 | |||
GPT Double Nickase Plasmid (h2) | sc-402909-NIC-2 | 20 µg | $410.00 |
Human GPT encodes glutamic-pyruvic transaminase (alanine aminotransferase 1), a pyridoxal phosphate–dependent enzyme that catalyzes reversible transamination between alanine and α-ketoglutarate to generate pyruvate and glutamate. This activity links amino acid catabolism to central carbon metabolism, supporting glycolysis/gluconeogenesis, nitrogen handling, and anaplerotic flux through the TCA cycle. GPT function contributes to metabolic reprogramming under nutrient stress and is frequently examined in contexts where amino acid utilization and mitochondrial-redox balance are altered, including metabolic dysfunction and cancer-associated metabolism.
GPT Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the GPT locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within GPT. 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 GPT 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 GPT-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.