Asparagine synthetase Double Nickase Plasmid (h): sc-402240-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Asparagine synthetase 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
- Asparagine synthetase Double Nickase Plasmid (h) and Asparagine synthetase Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ASNS. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Asparagine synthetase Antibody (G-10): sc-365809
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Asparagine synthetase Double Nickase Plasmid (h) | sc-402240-NIC | 20 µg | $410.00 | |||
Asparagine synthetase Double Nickase Plasmid (h2) | sc-402240-NIC-2 | 20 µg | $410.00 |
ASNS encodes human asparagine synthetase, a cytosolic amidotransferase that catalyzes ATP-dependent conversion of aspartate and glutamine to asparagine and glutamate. This activity links amino acid homeostasis to nitrogen metabolism and supports biosynthetic demands during nutrient stress through integration with the amino acid response and mTOR-regulated translational control. ASNS expression is dynamically regulated by cellular stress pathways such as ATF4 signaling, influencing adaptation to glutamine limitation and endoplasmic reticulum stress. Dysregulated ASNS activity and expression have been associated with altered metabolic states observed in cancer cell nutrient dependence and with neurodevelopmental disorders involving impaired asparagine biosynthesis.
Asparagine synthetase Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ASNS locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ASNS. 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 ASNS 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 ASNS-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.