ASCT2 Double Nickase Plasmid (m): sc-422987-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ASCT2 Double Nickase Plasmid (m) 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
- ASCT2 Double Nickase Plasmid (m) and ASCT2 Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Slc1a5. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ASCT2 Double Nickase Plasmid (m) | sc-422987-NIC | 20 µg | $410.00 |
Mouse Slc1a5 encodes the neutral amino acid transporter ASCT2 (SLC1A5), a sodium-dependent carrier that mediates cellular uptake and exchange of glutamine and other small neutral amino acids. ASCT2 supports amino acid homeostasis and fuels biosynthetic and bioenergetic demands by linking glutamine availability to pathways such as mTORC1 signaling, redox control via glutathione synthesis, and mitochondrial anaplerosis. In the immune system and rapidly proliferating cell types, ASCT2 activity influences activation programs and metabolic reprogramming that couple nutrient transport to growth and stress responses. Dysregulated SLC1A5 expression or function has been associated with altered nutrient dependency and metabolic phenotypes studied in cancer biology, inflammation, and tissue remodeling models.
ASCT2 Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Slc1a5 locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Slc1a5. 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 Slc1a5 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 Slc1a5-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.