CD73 Double Nickase Plasmid (h): sc-400307-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- CD73 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
- CD73 Double Nickase Plasmid (h) and CD73 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting NT5E. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: CD73 Antibody (D-12): sc-398260
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
CD73 Double Nickase Plasmid (h) | sc-400307-NIC | 20 µg | $410.00 | |||
CD73 Double Nickase Plasmid (h2) | sc-400307-NIC-2 | 20 µg | $410.00 |
Human NT5E encodes CD73 (ecto-5′-nucleotidase), a glycosylphosphatidylinositol-anchored surface enzyme that hydrolyzes extracellular AMP to adenosine, thereby shaping purinergic signaling and nucleotide balance in the pericellular space. By regulating adenosine accumulation, CD73 influences GPCR-mediated cAMP signaling, endothelial barrier function, leukocyte trafficking, and hypoxia-responsive programs that intersect with metabolic adaptation and inflammatory signaling. CD73 activity contributes to modulation of platelet and vascular responses and can impact epithelial–mesenchymal transition, stromal remodeling, and immune cell polarization in diverse tissue contexts. Dysregulated NT5E/CD73 expression or enzymatic activity has been associated with altered immune regulation, vascular calcification phenotypes, and tumor microenvironment biology, supporting its use as a mechanistic node in studies of inflammation, fibrosis, and cancer-associated pathways.
CD73 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the NT5E locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within NT5E. 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 NT5E 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 NT5E-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.