CPS2 Double Nickase Plasmid (h): sc-403519-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- CPS2 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
- CPS2 Double Nickase Plasmid (h) and CPS2 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting CAD. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: CPS2 Antibody (F-6): sc-376072
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
CPS2 Double Nickase Plasmid (h) | sc-403519-NIC | 20 µg | $410.00 | |||
CPS2 Double Nickase Plasmid (h2) | sc-403519-NIC-2 | 20 µg | $410.00 |
Human CAD encodes a multifunctional cytosolic enzyme that catalyzes the first three committed reactions of de novo pyrimidine biosynthesis, with the CPS2 domain mediating glutamine-dependent synthesis of carbamoyl phosphate. By coupling carbamoyl phosphate synthetase activity to aspartate transcarbamylase and dihydroorotase functions within a single polypeptide, CAD supports nucleotide production required for DNA replication, RNA transcription, and membrane phospholipid synthesis. CAD activity is coordinated with proliferative signaling and cellular metabolic state, linking pyrimidine availability to cell cycle progression and stress responses. Dysregulation of pyrimidine metabolism and altered CAD function have been associated with proliferative phenotypes and metabolic vulnerability in cancer-relevant models, making CAD a useful node for studying nucleotide homeostasis.
CPS2 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the CAD locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within CAD. 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 CAD 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 CAD-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.