AGAT Double Nickase Plasmid (h2): sc-405809-NIC-2
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- AGAT Double Nickase Plasmid (h2) 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
- AGAT Double Nickase Plasmid (h2) and AGAT Double Nickase Plasmid (h22) encode distinct paired gRNA designs targeting GATM. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
AGAT Double Nickase Plasmid (h2) | sc-405809-NIC-2 | 20 µg | $410.00 |
Human GATM encodes L-arginine:glycine amidinotransferase (AGAT), a mitochondrial enzyme that catalyzes the first committed step of creatine biosynthesis by converting arginine and glycine to guanidinoacetate, thereby supporting cellular energy buffering through the creatine–phosphocreatine system. AGAT activity links amino acid metabolism with mitochondrial function and bioenergetic pathways, influencing tissues with high ATP demand and intersecting with nitric oxide and one-carbon metabolic networks via substrate availability. Genetic disruption of GATM is associated with creatine deficiency and neurometabolic phenotypes, and altered expression has been investigated in renal and muscle physiology as well as broader metabolic stress responses. Gene editing of GATM in human cell models enables mechanistic studies of creatine pathway regulation, mitochondrial metabolism, and genotype–phenotype relationships, including validation of variants and development of functional assays for metabolic flux and energy homeostasis.
AGAT Double Nickase Plasmid (h2) consists of a matched pair of plasmids engineered for high-specificity editing of the GATM locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within GATM. 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 GATM 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 GATM-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.