AdoMetDC Double Nickase Plasmid (h): sc-404514-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- AdoMetDC 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
- AdoMetDC Double Nickase Plasmid (h) and AdoMetDC Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting AMD1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: AdoMetDC Antibody (E-6): sc-166970
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
AdoMetDC Double Nickase Plasmid (h) | sc-404514-NIC | 20 µg | $410.00 | |||
AdoMetDC Double Nickase Plasmid (h2) | sc-404514-NIC-2 | 20 µg | $410.00 |
AMD1 encodes human S-adenosylmethionine decarboxylase (AdoMetDC), a rate-limiting enzyme in polyamine biosynthesis that generates decarboxylated S-adenosylmethionine used for spermidine and spermine production. Through regulation of intracellular polyamine pools, AdoMetDC influences translation, chromatin organization, DNA replication, and cell-cycle progression, linking AMD1 activity to growth control and cellular stress responses. Dysregulated polyamine metabolism and altered AMD1 expression or activity have been associated with proliferative phenotypes and metabolic reprogramming observed across multiple disease contexts, supporting its use as a mechanistic node for pathway interrogation. As a metabolic gatekeeper, AMD1 is commonly studied in connection with ornithine decarboxylase and polyamine catabolism/transport pathways to map homeostatic feedback and downstream gene-expression effects.
AdoMetDC Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the AMD1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within AMD1. 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 AMD1 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 AMD1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.