Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Acid Ceramidase Double Nickase Plasmid (h): sc-401495-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Acid Ceramidase 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
- Acid Ceramidase Double Nickase Plasmid (h) and Acid Ceramidase Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ASAH1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Acid Ceramidase Antibody (23): sc-136275
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Acid Ceramidase Double Nickase Plasmid (h) | sc-401495-NIC | 20 µg | $410.00 | |||
Acid Ceramidase Double Nickase Plasmid (h2) | sc-401495-NIC-2 | 20 µg | $410.00 |
ASAH1 encodes acid ceramidase, a lysosomal hydrolase that cleaves ceramide into sphingosine and free fatty acid, linking sphingolipid catabolism to sphingosine-1-phosphate signaling. By regulating the ceramide–sphingosine rheostat, acid ceramidase influences membrane turnover, lysosomal homeostasis, autophagy, and stress-responsive pathways that shape cell survival and inflammatory signaling. Dysregulated ASAH1 activity perturbs sphingolipid balance and has been associated with lysosomal storage pathology and neurodegenerative phenotypes, as well as altered lipid signaling in cancer and metabolic stress models. ASAH1 is therefore widely studied in the context of lipid-mediated signaling, organelle quality control, and sphingolipid-driven modulation of cellular fate decisions.
Acid Ceramidase Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ASAH1 locus in human 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.