Acid Ceramidase Double Nickase Plasmid (m): sc-419216-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Acid Ceramidase 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
- Acid Ceramidase Double Nickase Plasmid (m) and Acid Ceramidase Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Asah1. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Acid Ceramidase Double Nickase Plasmid (m) | sc-419216-NIC | 20 µg | $410.00 | |||
Acid Ceramidase Double Nickase Plasmid (m2) | sc-419216-NIC-2 | 20 µg | $410.00 |
Mouse Asah1 encodes acid ceramidase, a lysosomal hydrolase that deacylates ceramide to sphingosine and free fatty acids, thereby controlling the balance between ceramide and sphingosine-1-phosphate signaling. Through regulation of sphingolipid turnover, Acid Ceramidase influences membrane composition, endolysosomal homeostasis, autophagy, and stress-response pathways that intersect with apoptosis and inflammatory signaling. Disruption of ASAH1-dependent ceramide catabolism perturbs lysosomal lipid clearance and alters bioactive sphingolipid pools, processes implicated in neurodegeneration and systemic lipid storage phenotypes. Asah1 is therefore a key node for studying lysosomal function and sphingolipid-driven signaling networks in mouse cellular and in vivo models.
Acid Ceramidase Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Asah1 locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Asah1. 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 Asah1 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 Asah1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.