AANAT Double Nickase Plasmid (h): sc-403080-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- AANAT 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
- AANAT Double Nickase Plasmid (h) and AANAT Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting AANAT. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
AANAT Double Nickase Plasmid (h) | sc-403080-NIC | 20 µg | $410.00 | |||
AANAT Double Nickase Plasmid (h2) | sc-403080-NIC-2 | 20 µg | $410.00 |
Human AANAT (arylalkylamine N-acetyltransferase) is the rate-limiting enzyme in melatonin biosynthesis, catalyzing acetylation of serotonin to N-acetylserotonin within the tryptophan/indoleamine metabolic pathway. Its activity is tightly regulated by circadian and cAMP/PKA signaling, including phosphorylation-dependent stabilization and interactions that coordinate nocturnal melatonin production. Through control of melatonin and related indoleamine intermediates, AANAT contributes to sleep–wake and neuroendocrine timing, oxidative stress responses, and metabolic homeostasis. Altered AANAT expression or regulation has been investigated in circadian rhythm disruption and sleep-related phenotypes, as well as neuropsychiatric and metabolic research contexts.
AANAT Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the AANAT locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within AANAT. 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 AANAT 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 AANAT-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.