dUTPase Double Nickase Plasmid (h): sc-405843-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- dUTPase 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
- dUTPase Double Nickase Plasmid (h) and dUTPase Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting DUT. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: dUTPase Antibody (H-9): sc-166856
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
dUTPase Double Nickase Plasmid (h) | sc-405843-NIC | 20 µg | $410.00 | |||
dUTPase Double Nickase Plasmid (h2) | sc-405843-NIC-2 | 20 µg | $410.00 |
DUT encodes human dUTPase, a nucleotide-metabolizing enzyme that hydrolyzes dUTP to dUMP, thereby lowering intracellular dUTP pools and supplying substrate for thymidylate biosynthesis. By preventing uracil misincorporation into DNA, dUTPase supports genome integrity during DNA replication and repair and helps limit base-excision repair cycles triggered by uracil-DNA glycosylases. DUT activity integrates with folate-dependent one-carbon metabolism and pyrimidine homeostasis, linking nucleotide balance to replication stress responses. Dysregulated dUTP metabolism and uracil accumulation are associated with elevated DNA damage signaling and have been investigated in contexts of genomic instability and tumor biology.
dUTPase Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the DUT locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within DUT. 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 DUT 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 DUT-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.