ADO Double Nickase Plasmid (h): sc-408985-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ADO 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
- ADO Double Nickase Plasmid (h) and ADO Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ADO. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: ADO Antibody (E-11): sc-515318
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ADO Double Nickase Plasmid (h) | sc-408985-NIC | 20 µg | $410.00 | |||
ADO Double Nickase Plasmid (h2) | sc-408985-NIC-2 | 20 µg | $410.00 |
Human ADO (2-aminoethanethiol dioxygenase) is a non-heme iron enzyme that catalyzes the oxygenation of cysteamine to hypotaurine, linking coenzyme A turnover to taurine biosynthesis and cellular redox balance. By controlling the flux of reactive aminothiols, ADO influences thiol homeostasis, antioxidant capacity, and metabolic adaptation to oxidative and nutrient stress. The ADO-dependent cysteamine–hypotaurine axis intersects with sulfur amino acid metabolism and can modulate downstream processes such as mitochondrial function and inflammatory signaling. Altered aminothiol metabolism and taurine availability have been associated with cardiometabolic and neurobiological phenotypes, making ADO a useful target for mechanistic studies of stress-responsive metabolism.
ADO Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ADO locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ADO. 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 ADO 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 ADO-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.