Aspartoacylase Double Nickase Plasmid (h): sc-405771-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Aspartoacylase 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
- Aspartoacylase Double Nickase Plasmid (h) and Aspartoacylase Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ASPA. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Aspartoacylase Antibody (D-11): sc-377308
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Aspartoacylase Double Nickase Plasmid (h) | sc-405771-NIC | 20 µg | $410.00 | |||
Aspartoacylase Double Nickase Plasmid (h2) | sc-405771-NIC-2 | 20 µg | $410.00 |
Human ASPA encodes aspartoacylase, a zinc-dependent hydrolase that deacetylates N-acetyl-L-aspartate (NAA) to generate acetate and L-aspartate, supporting carbon flux for lipid synthesis and neuronal–glial metabolic coupling in the central nervous system. By regulating NAA turnover, ASPA contributes to myelin-related processes and acetate availability used in acetyl-CoA-dependent pathways, including membrane biogenesis and protein acetylation. Loss-of-function variants in ASPA are linked to Canavan disease, a leukodystrophy characterized by abnormal NAA accumulation and dysmyelination, making ASPA a key target for studying neurodevelopmental metabolism and glial biology. ASPA perturbation models are frequently used to interrogate metabolite homeostasis, oligodendrocyte function, and downstream transcriptional and bioenergetic adaptations.
Aspartoacylase Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ASPA locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ASPA. 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 ASPA 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 ASPA-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.