AATC Double Nickase Plasmid (h): sc-405477-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- AATC 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
- AATC Double Nickase Plasmid (h) and AATC Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting GOT1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: AATC Antibody (H-8): sc-515641
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
AATC Double Nickase Plasmid (h) | sc-405477-NIC | 20 µg | $410.00 |
Human GOT1 encodes cytosolic aspartate aminotransferase (AATC), a pyridoxal phosphate–dependent enzyme that catalyzes reversible transamination between aspartate and α-ketoglutarate to generate oxaloacetate and glutamate. This activity links amino acid metabolism to the malate–aspartate shuttle, supporting NADH redox balance, anaplerosis, and carbon flux into the TCA cycle. GOT1 function interfaces with nitrogen handling and glutamate/aspartate pools that influence biosynthetic programs and cellular stress responses. Dysregulation of GOT1-associated metabolic routing has been studied in contexts of altered bioenergetics and redox homeostasis relevant to proliferative and degenerative disease mechanisms.
AATC Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the GOT1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within GOT1. 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 GOT1 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 GOT1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.