AATM Double Nickase Plasmid (h): sc-403969-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- AATM 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
- AATM Double Nickase Plasmid (h) and AATM Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting GOT2. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: AATM Antibody (E-7): sc-271702
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
AATM Double Nickase Plasmid (h) | sc-403969-NIC | 20 µg | $410.00 | |||
AATM Double Nickase Plasmid (h2) | sc-403969-NIC-2 | 20 µg | $410.00 |
Human GOT2 encodes mitochondrial aspartate aminotransferase (AATM), a pyridoxal phosphate–dependent enzyme that catalyzes the reversible transamination between glutamate/oxaloacetate and α-ketoglutarate/aspartate. This activity is central to the malate–aspartate shuttle, coupling cytosolic and mitochondrial redox balance while supplying aspartate for nucleotide biosynthesis and supporting anaplerotic flux into the TCA cycle. Through these pathways, AATM links amino acid metabolism to oxidative phosphorylation and cellular proliferation programs. Dysregulated GOT2/AATM function has been investigated in contexts of metabolic rewiring and mitochondrial dysfunction associated with cancer biology and other disorders of intermediary metabolism.
AATM Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the GOT2 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within GOT2. 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 GOT2 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 GOT2-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.