V-ATPase A1 Double Nickase Plasmid (h): sc-400975-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- V-ATPase A1 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
- V-ATPase A1 Double Nickase Plasmid (h) and V-ATPase A1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ATP6V0A1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: V-ATPase A1 Antibody (E-8): sc-374475
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
V-ATPase A1 Double Nickase Plasmid (h) | sc-400975-NIC | 20 µg | $410.00 | |||
V-ATPase A1 Double Nickase Plasmid (h2) | sc-400975-NIC-2 | 20 µg | $410.00 |
ATP6V0A1 encodes the a1 isoform of the V0 domain a subunit within vacuolar H\+-ATPase (V-ATPase), a multi-subunit proton pump that acidifies endosomes, lysosomes, and secretory vesicles. V-ATPase-driven organelle acidification supports receptor-mediated endocytosis, lysosomal degradation, autophagy, and vesicular trafficking, and it influences mTORC1 signaling and cellular nutrient sensing through lysosome function. Through control of luminal pH, V-ATPase A1 also contributes to proteostasis and membrane fusion events required for efficient cargo sorting. Dysregulation of V-ATPase activity and endolysosomal acidification has been linked to neurodevelopmental and neurodegenerative phenotypes, altered synaptic function, and broader defects in cellular homeostasis relevant to disease mechanisms.
V-ATPase A1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ATP6V0A1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ATP6V0A1. 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 ATP6V0A1 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 ATP6V0A1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.