ASM Double Nickase Plasmid (h): sc-401347-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ASM 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
- ASM Double Nickase Plasmid (h) and ASM Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting SMPD1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: ASM Antibody (4H2): sc-293189
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ASM Double Nickase Plasmid (h) | sc-401347-NIC | 20 µg | $410.00 | |||
ASM Double Nickase Plasmid (h2) | sc-401347-NIC-2 | 20 µg | $410.00 |
SMPD1 encodes acid sphingomyelinase (ASM), a lysosomal hydrolase that converts sphingomyelin to ceramide and phosphocholine, thereby influencing sphingolipid turnover and membrane lipid composition. ASM-dependent ceramide production contributes to endolysosomal trafficking, autophagy-lysosome function, stress-responsive signaling, and organization of membrane microdomains that modulate receptor signaling. Disruption of SMPD1 alters sphingomyelin/ceramide balance and impacts inflammatory and apoptotic pathways linked to cellular homeostasis. Genetic defects in SMPD1 are associated with lysosomal storage pathology consistent with Niemann–Pick disease types A/B, making SMPD1 a widely used target in studies of lysosomal biology and sphingolipid metabolism.
ASM Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the SMPD1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within SMPD1. 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 SMPD1 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 SMPD1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.