ITPase Double Nickase Plasmid (h): sc-410841-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ITPase 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
- ITPase Double Nickase Plasmid (h) and ITPase Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ITPA. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: ITPase Antibody (A-4): sc-514409
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ITPase Double Nickase Plasmid (h) | sc-410841-NIC | 20 µg | $410.00 |
Human ITPA encodes inosine triphosphate pyrophosphatase (ITPase), a nucleotide pool–sanitizing enzyme that hydrolyzes noncanonical purine triphosphates such as ITP and dITP to prevent their misincorporation into RNA and DNA. By limiting inosine-containing nucleic acids, ITPase supports replication fidelity, transcriptional integrity, and broader genome maintenance processes linked to base excision repair and replication stress responses. Perturbation of ITPA activity alters purine metabolism and can increase sensitivity to imbalanced nucleotide pools, with downstream effects on mitochondrial and nuclear DNA stability. Genetic variation or reduced ITPase function has been associated with susceptibility to nucleic-acid damage phenotypes and differential responses in studies of purine analog metabolism, making it relevant for mechanistic investigations of genome integrity.
ITPase Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ITPA locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ITPA. 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 ITPA 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 ITPA-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.