ASL Double Nickase Plasmid (h): sc-403083-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ASL 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
- ASL Double Nickase Plasmid (h) and ASL Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ASL. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: ASL Antibody (B-1): sc-166787
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ASL Double Nickase Plasmid (h) | sc-403083-NIC | 20 µg | $410.00 | |||
ASL Double Nickase Plasmid (h2) | sc-403083-NIC-2 | 20 µg | $410.00 |
Human ASL encodes argininosuccinate lyase, a cytosolic enzyme that catalyzes the reversible cleavage of argininosuccinate to arginine and fumarate, linking the urea cycle to cellular energy metabolism via the TCA cycle. By supporting nitrogen disposal and arginine availability, ASL influences amino acid homeostasis and downstream nitric oxide biology through effects on arginine supply. ASL activity is central to hepatic ammonia detoxification and broader metabolic coordination across tissues. Disruption of ASL function is associated with urea cycle dysfunction and hyperammonemia-related metabolic phenotypes, making it relevant for mechanistic studies of nitrogen metabolism and mitochondrial–cytosolic crosstalk.
ASL Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ASL locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ASL. 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 ASL 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 ASL-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.