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.
NPC1 Double Nickase Plasmid (m): sc-421941-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- NPC1 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
- NPC1 Double Nickase Plasmid (m) and NPC1 Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Npc1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: NPC1 Antibody (G-1): sc-271335
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
NPC1 Double Nickase Plasmid (m) | sc-421941-NIC | 20 µg | $410.00 | |||
NPC1 Double Nickase Plasmid (m2) | sc-421941-NIC-2 | 20 µg | $410.00 |
Mouse Npc1 encodes NPC1, a multi-pass lysosomal membrane protein that cooperates with NPC2 to mobilize unesterified cholesterol and other lipids from late endosomes/lysosomes to cellular membranes. By governing intracellular sterol trafficking, NPC1 influences cholesterol homeostasis, lysosome function, and downstream metabolic signaling pathways that depend on proper membrane lipid composition. Disruption of NPC1 activity leads to lysosomal lipid accumulation and broad perturbations in endolysosomal transport and autophagy-associated processes. Npc1 is therefore widely used as a genetic entry point to model lipid storage phenotypes and to dissect mechanisms of lysosome-centered metabolic regulation in mouse cells.
NPC1 Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Npc1 locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Npc1. 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 Npc1 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 Npc1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.